life science archives (lsa)by dr. rosukon can be classify as a complex organic substance of protein...

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Life Science Archives (LSA)

ISSN: 2454-1354

Volume – 4; Issue - 2; Year – 2018; Page: 1292 – 1308

DOI: 10.22192/lsa.2018.4.2.1

©2018 Published by JPS Scientific Publications Ltd. All Rights Reserved

Review Article

AN EXCLUSIVE REVIEW ON PRODUCTION, ANALYSIS AND APPLICATIONS

OF ENZYME BIO-CLEANERS FROM FRUIT AND VEGETABLE WASTE

Ashwani Kumar*1, Vaishali Dhiman

2, Anuradha Saini

2 and Amit Panwar

2

1*Dean, Faculty of Biosciences, Shri Ram College Muzaffarnagar, Uttar Pradesh – 251 001, India, 2PG Student of Biosciences, Shri Ram College Muzaffarnagar, Uttar Pradesh - 251001, India,

Abstract

The waste generated from fruits and vegetables are organic in nature and supply a major share in pollution of air, water and soil. Fruit and vegetable wastes (FVWs) is a matter of serious environmental

concern due to emission of green house gases. Enzyme Bio-cleaners are the an organic compounds including

enzymes produced by the simple fermentation of fresh vegetable wastes, fruit wastes with addition of brown

sugar and water by using the selective microorganisms like Yeast and Bacteria. This anaerobic fermentation

generates natural chains of proteins, minerals, salts, organic acids, alcohol and enzymes, which have the

capacity to breakdown, modify, create and catalyze - functions that make it an astonishing cleaning support

in household as well as other applications. The target enhancement plan at enzymes with enhanced stability,

improved specific activity, modification of pH-activity profiles, and increased catalytic specificity will be the

main focus in future research. This has been mostly achieved through refined fermentation techniques, along

with improvements in screening procedures. To dispose of dirt and soils safely, economically and rapidly

and digest chemical and organic waste without odour or harmful gas the enzyme bio-cleaning solutions

formulated instead of noxious chemicals. So it is utmost important to develop the best suitable alternate to

the synthetic cleaning solution with increased efficiency and stability in different environmental conditions.

This review deals with the developments on microbial processing technologies for production of useful

enzymes from FVWs. The incorporation of genetic engineering for improvement of microbial strains in

order to enhance the production with value added bio-based products as well as the concept of zero-waste

economy are the key parameters for enzyme bio-cleaners. In this framework, enzyme bio-cleaners referred to

as natural green chemicals are becoming a perfect future consumer preference for miscellaneous applications

and sustainable environment.

Article History Received : 05.01.2018

Revised : 08.02.2018

Accepted: 15.03.2018

Key words: Fruit and vegetable wastes, Enzyme,

Bio-cleaners, Anaerobic fermentation.

1. Introduction

In India, Fruit and Vegetable Wastes

constitute about more than 10 million tons

annually and currently these wastes are disposed

* Corresponding author: Ashwani Kumar E.mail: [email protected]

by dumping on the outskirts of cities. The main

contributors of waste generation in Indian society

are agriculture and municipal sectors. Industrial

sectors have also not been able to handle and treat

the wastes generated by them or control the

mission of obnoxious gases into the atmosphere.

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Ashwani Kumar/Life Science Archives (LSA), Volume – 4, Issue – 2, 2018, Page – 1292 to 1308 1293


In spite of the various environmental rules and

regulations, very little has been achieved in terms

of minimization of waste generations. In Indian

cities total quantum of waste generation is

increasing at rate of 1.33 % (Srilatha et al., 1995).

Enzymes are proteins, which act as

catalysts. Enzymes lower the energy required for

are action to occur, without being used up in the

reaction. Many types of industries, to aid in the

generation of their products, utilize enzymes.

Enzymatic processes have been implemented in a

broad range of industries in recent decades

because they are specific, fast in action and often

save raw materials, energy, chemicals and water

compared to conventional processes. A number of

comparative environmental assessment studies

have been conducted in the past fifteen years to

investigate whether these properties of enzymatic

processes lead to environmental improvements

and assess whether they could play a role in

moving toward cleaner industrial production. On

the other hand, enzymes have become popular in

cleaning products because they are biodegradable

and facilitate selective reactions. Fermentation is a

method of generating enzymes for industrial

purposes. Fermentation involves the use of

microorganisms, like bacteria and yeast to produce

the enzymes. Enzyme Bio cleaners are an organic

solution produced by the simple fermentation of

fresh vegetable wastes, fruit wastes with addition

of brown sugar and water by using the selective

microorganisms like Yeast and Bacteria. This

fermentation creates natural chains of proteins,

mineral salts, organic acids, alcohol and enzymes.

This solution has the capacity to breakdown,

change, create and catalyse functions that make it

a wonderful cleaning aid in household as well as

in industrial and medical applications

(Thirumurugan and Mathivanan, 2016).

Biocleaning enzyme solution was

developed in Thailand through simple inexpensive

method for various household and hospital

cleaning methods. This has been developed for

people to make simple cleaning solution at home

or in small scale business to ease global warming

because minimizing the environmental pollution

by utilizing the garbage like kitchen waste

vegetable and fruit wastes from stalls and markets.

This product is natural, safe, and good for the

environment and uses the magic of nature to

produce extraordinary removal of dirt; oil stain

etc. resulted to customer satisfaction. During the

past few years, there has been serious public

concern about the ecological problems arising

from the use of synthetic cleaning agent which

releases toxic chemicals and unstable pH and

chemicals of environment. Owing to the fact that

these synthetic cleaning solutions are corrosive,

toxic and exhibit a slow rate of biodegradation,

their extensive usage leads to the formation of

slumps, creating unhygienic conditions in the

surroundings However, the latest cleaning

technologies include enzyme containing cleaning

solutions are very effective, safe and specificity in

activity and environmental friendly as well as

cheaper preparation methods. Enzyme Bio-

Cleaning solution is formulated specifically to

dispose of dirt and soils safely, economically and

rapidly and work quickly and efficiently to digest

chemical and organic waste with no odour or

noxious gas. So suitable alternate to the synthetic

cleaning solution with regard to biodegradability,

low toxicity, non-Corrosiveness environment-

friendliness, enhanced cleaning properties as well

as increased efficiency and stability in different

environmental conditions are require to develop

(Thirumurugan and Mathivanan, 2016).

In this context, enzyme bio-cleaners

referred to as green chemicals are becoming an

ideal consumer choice for various applications.

The advantage of using enzyme bio-cleaning

solution are 1) It is safer for the environment and

safer for human health than traditional chemical

cleaners and odour control products, 2) It is highly

specialized enzyme producing microorganisms to

clean and control odours by eliminating the soils

rapidly, 3) It is economically cheaper and the cost

of production is less 4) This Bio-Cleaning solution

provide residual cleaning for longer period and

gives stable application and 5) This Enzyme Bio

cleaning solution help to displace unknown,

potentially pathogenic (disease causing) bacteria

with known, healthy microorganisms and in this

way contribute to better human health. Thus to

conclude, cleaning forms an important aspect for



the maintenance of hygiene and safety of foods in

the food processing industry. Due to their high

efficiency and safety, it is assumed that the

enzyme cleaners will eventually capture a bulk of

the Indian market. Some uses recommended for

this non toxic, environmentally friendly enzyme:

1) For dishes and laundry. 2) For washing

bathrooms and toilets, grime comes off easily. 3)

For removing stubborn stains and odours

(coloured fabrics and floors). 4) To clean

vegetables and fruits. 5) Clear blockages in

kitchen sinks and drains (use blended pulp/sludge

of enzyme) 6) As a repellent use undiluted) for

ants, and cockroaches. 7) Form mopping floors8)

As fertilizer for plants in garden. 9) As a skincare

product, e.g. facial cleanser or toner. 10) Wash

cars. 11) Save Money, Save Space, Save Water

and Save The Earth! the above scientific and

technical information, the present work is

proposed to carry out the production and analysis

of the Enzyme Bio-cleaning solution by using the

residues and waste of agricultural produces like

fruit and kitchen waste including vegetable wastes

with help of Yeast (Sacharomyces sp.) and

Bacteria (Bacillus sp.) with addition of cheaper

carbohydrate sources like brown sugar and water

medium. Garbage Enzyme Since there are many

drawbacks of the lake restoration in terms of

pollution control an alternative method was

studied to treat the water pollution problems. This

method involves the use of garbage enzyme which

is produced from the food waste such as fruit

peeled through fermentation with brown sugar for

3 months time. Then, after 3 months the enzymes

are ready to use as a household cleaning liquid, to

remove foul odours, toilet, anti-bacterial and anti-

viral agent. This garbage enzyme which invented

by Dr. Rosukon can be classify as a complex

organic substance of protein chains and mineral

salts and juvenile hormones. Researchers

postulated that isozyme can functions in four

categories: decompose, compose, transform and

catalysis (Thirumurugan and Mathivanan, 2016).

Figure – 1: Base flow of kitchen waste

management

Intravascular thrombosis is one of the

major causes of variety of cardiovascular disorders

leading to high mortality worldwide.

Fibrinolytic enzymes from microbial sources

possess ability to dissolve these clots and help to

circumvent these problems in more efficient and

safer way. The purified enzyme belongs to metallo

protease class and had optimal activity in

conditions similar to physiological environment

with temperature optima of 40 °C and pH optima

of Roohi et al. (2017). The optimized parameters

under solid-state fermentation, lipase and phytase

activities were recorded as 1056.66±2.92 and

833±2.5 U/g of substrate (U/g), respectively.

Besides enzyme production, complete elimination

of phorbol esters and significant phytate reduction

from 6.51±0.01 to 0.43±0.01g/100 g of seed cake

were noted after 3days incubation. Curcin and

trypsin inhibition activity were reduced

significantly from 26.33±0.43 to 0.56±0.02

mg/100 g and 229.33±2.02 to 11.66±0.28 U/g,

respectively after 5 days incubation. Saponins

were reduced from 5.56±0.19 to 1.95±0.01 g/100

g of seed cake after 7 days incubation (Kannoju et

al., 2017). In this study, we used multiple

metabolomic approaches to characterize the

microbial community and the active metabolic

pathways of a stable industrial biogas reactor with

food waste as the dominant feedstock, operating at



thermophilic temperatures (60 °C) and elevated

levels of free ammonia (367 mg/liter NH3-N). The

microbial community was strongly dominated (76

% of all 16S rRNA amplicon sequences) by

populations closely related to the proteolytic

bacterium Coprothermobacter proteolyticus. We

discuss the impact of frequently encountered

microbial populations as well as the metabolism of

newly discovered novel phylotypes that seem to

play distinct roles within key microbial stages of

anaerobic digestion in this stable high-temperature

system. In particular, we draft a metabolic

scenario whereby multiple uncultured syntrophic

acetate-oxidizing bacteria are capable of

syntrophically oxidizing acetate as well as longer-

chain fatty acids (via the β-oxidation and Wood-

Ljundahl pathways) to hydrogen and carbon

dioxide, which methanogens subsequently convert

to methane (Hagen et al., 2016).

Ectomycorrhizal fungi are an essential

component of forest ecosystems, most of which

can form edible and medical fruiting bodies. The

fructification and hypogeous community structure

of ectomy corrhizal fungi symbiosing with cedar

(Cedrus deodara (Roxb.) G. Don). The results

showed that garbage significantly altered soil

abiotic and biotic properties, increasing soil urease

activity, decreasing the soil exchangeable metal

content and phos phatase activity, and ultimately

inhibiting the formation of fruiting bodies. The

pollution of garbage also changed the community

structure of hypo geous ectomy corrhizal fungi

where ectomy corrhizal ascomycetes dominated.

In unpolluted sites, the relative abundance of

ectom ycorrhizal ascomcetes and basidiomycetes

were almost equal. Although, no fruiting bodies

were observed in that soil polluted by

living garbage, the sequencing result showed that

various ectomy corrhizal fungi were present

underground, suggesting that these taxonomic

fungi had the potential to cope with adverse

conditions. This study not only provided a deeper

understanding of the relationship between ectomy

corrhizal fungal communities and prevailing

environmental conditions, but provided a new

pathway for the excavation and utilization of the

resource of antistress ectomy corrhizal fungi (Sun

et al., 2017). The production of carboxy methyl

cellulase (CMCase) by Bacillus halodurans

IND18 under Solid Substrate Fermentation (SSF)

using cow dung was optimized through two level

full factorial design and second order Response

Surface Methodology (RSM). The central

composite design (CCD) was employed to

optimize the vital fermentation parameters, such

as pH of the substrate, concentration of nitrogen

source (peptone) and ion (sodium dihydrogen

phosphate) sources in medium for achieving

higher enzyme production. This study

demonstrated the potential of cow dung as novel

substrate for CMCase production (Vijayaraghavan

et al. 2016).

Use black sugar, brown or any raw unprocessedSugar but never use white sugar

Use kitchen waste/ fruit skins like apple, orange, papaya,

banana, carrot but don’t use durians! Never use meat!

Add sugar + fruit skins +water and mix well in a plastic bottle and cap tightly

Keep in a cool dry place at room temperature. Release the gases formed once a day for the first month. Make sure it is re-capped tightly

For the 2nd and 3rd months, only release any gases if necessary

Fermentation will take place in the bottles for 3 months. These bottles contain kitchen waste with sugar and water

Then after filtration and removal of the residue, garbage enzyme is obtained

Figure – 2: Production flow sheet of enzyme

bio-cleaner



GARBAGE ENZYME’S

MANY USES

Cleaning the house

Cleaning the toilet

Cleaning the kitchen

Cleaning vegetables

Cleaning clothes

Garden care

Skin care

Car care

Pet care

Purify air

Deodorize air

Fig – 3: Applications of Garbage enzyme bio-

cleaner

2. Fermented products from fruit waste

The importance of soil fungi in complex

carbon degradation and the recent identification of

genes involved in this process have sparked

considerable interest in examining fungal gene

expression in situ. In sum, the reduced richness

and altered composition produced in the d s cDNA

libraries could substantially influence ecological

interpretations of the data. Defining the factors

underpinning the methodological biases will

potentially aid in optimizing the design of gene

expression studies in soils and other complex

environmental samples (Weber and Kuske, 2011).

A two-stage fermentation process, consisting of a

simultaneous saccharification and fermentation

(SSF) stage and a dry methane fermentation stage,

was developed to utilize garbage for the

production of fuel ethanol and methane.

Approximately, 85 % of the energy of

the garbage was converted to fuels, ethanol and

methane by this process (Koike et al., 2009).

Unwanted or misfolded proteins are either

refolded by chaperones or degraded by the

ubiquitin-proteasome system (UPS ITC-induced

ALS is a proteasome-dependent assembly for

emergent removal of misfolded proteins,

suggesting that the cell may have a previously

unknown strategy to cope with misfolded proteins

(Mi et al., 2009). East Kolkata Wetlands is a

conserved wetland utilizing sewage and garbage,

generated by Kolkata Municipal Corporation area

for cultivation purpose No DNA damage was

monitored upon induction with Pb(2+) (Roy et al.,

2008). Thermophilic poly-L-lactide-degrading

bacteria were isolated from a garbage fermentor.

One of the isolates, strain PL21, was identified as

Bacillus smithii based on its physiological

properties, sugar assimilation pattern, and partial

16S rDNA sequence (Kim et al., 2005).

The

ubiquitin-mediated proteolysis pathway has come

a long way in the past decade. I focus particularly

on the known regulatory enzymes, and also on the

housekeeping enzymes that are implicated in

development of disease (Liu and Ge, 2009).

Figure – 4: Production of ethanol from fruit

waste

3. Fruit waste problem management and

prospects

Urbanization and industrialization

accompanied by population flare-up has formed a

serious problem of waste generation and its

disposal, treatment and management. Fruit and

vegetable wastes are produced in large quantities

in markets and constitute a source of nuisance in

municipal landfills because of their high (Viturtia

et al., 1989). In order to sustain a future bio-based

economy, methods for the conversion of

renewable feed stocks into the respective value-

added products will need to be efficient.

Furthermore, there is a need to explore all

possibilities in the use of sustainable resources to

ensure the extraction of maximum value with

minimum negative impact (Gama et al., 2013). In

this work, a low-cost alternative approach (i.e.,



adding aged refuse (AR) into waste activated

sludge) to significantly enhance anaerobic

digestion of sludge was reported. Experimental

results showed that with the addition dosage of

AR increasing from 0 to 400 mg/g dry sludge

soluble chemical oxygen demand (COD)

increased from 1150 to 5240 mg/L at the digestion

time of 5 d, while the maximal production of

volatile fatty acids (VFA) increased from 82.6 to

183.9 mg COD/g volatile suspended solids.

Although further increase of AR addition

decreased the concentrations of both soluble COD

and VFA, their contents in these systems with AR

addition at any concentration investigated were

still higher than those in the blank, which resulted

in higher methane yields in these systems. The

effect of heavy metals in AR on sludge anaerobic

digestion was dosage dependent. Sludge anaerobic

digestion was enhanced by appropriate amounts of

heavy metals but inhibited by excessive amounts

of heavy metals. The relative abundances of

microorganisms responsible for sludge hydrolysis

and acidogenesis were also observed to be

improved in the system with AR addition, which

was consistent with the performance of anaerobic

digestion (Taneja et al., 2017).

Food waste is a

sustainable feedstock that can be converted into

ethanol without an expensive thermo chemical

pretreatment that is commonly used in first and

second generation processes.

Optimized enzyme cocktail was found to

be 45% alpha amylase, 45 % gamma amylase, and

10 % pectinase at 2.5 mg enzyme protein/g glucan

produced a hydrolysate with high glucose

concentration. All three solid loadings (20 %, 30

%, and 40 %) produced sugar-rich hydrolysates

and ethanol with little to no enzyme or yeast

inhibition. Enzymatic hydrolysis and fermentation

process mass balance was carried out using pie

waste on a 1000 g dry weight basis that produced

329 g ethanol at 20 % solids loading. This process

clearly demonstrates how food waste could be

efficiently converted to ethanol that could be used

for making biodiesel by reacting with waste

cooking oil (Arun and Sivashanmugam, 2015).

Reuse and management of organic solid

waste, reduce the environmental impact on human

health and increase the economic status by

generating valuable products for current and novel

applications. Garbage enzyme is one such product

produced from fermentation of organic solid waste

and it can be used as liquid fertilizer, antimicrobial

agents, treatment of domestic wastewater,

municipal and industrial sludge treatment, etc This

result confirms matured garbage enzyme contains

a higher concentration of acetic acid and thus it

can be used as a monitoring parameter for semi-

continuous production of garbage enzyme in large

scale (Karmee et al., 2015). Depletion of fossil

fuels and environmental problems are encouraging

research on alternative fuels of renewable sources.

However, the cost of biodiesel production is high

and is attributed mainly to the feedstock used

which leads to the investigation of low cost

feedstock‟s that are economically feasible. The

maximum biodiesel yield was 100 % for the base

(KOH) catalyzed transesterification at 1:10M ratio

of lipid to methanol in 2 hrs at 60 °C. Novozyme-

435 yielded a 90 % FAME conversion at 40 °C

and 1:5 lipid to methanol molar ratio in 24 hrs.

Lipid obtained from fungal hydrolysis of food

waste is found to be a suitable feedstock for

biodiesel production (Hailei et al., 2015). A strain

of Ganoderma lucidum was separated and

identified according to its morphological

characteristics and phylogenetic data In 15 l

bioreactor, FW is also suitable for laccase

production and the maximum laccase activity

reached 54,000 U/l. Moreover, some details of

laccase over production using FW were

investigated. The G. lucidum consumes FW by

secreting a series of hydrolases and proteases and

the improvement of laccase activity is because FW

induces over-expression of three isoenzymes by

poly acrylamide gel electrophoresis analysis

(Chang et al., 2013).

The composting procedure in food waste

plants generates airborne bioaerosols that have the

potential to damage human airway epithelial cells.

Persistent inflammation and repair responses

induce airway remodeling and damage to the

respiratory system. This study elucidated the

expression changes of airway remodeling genes in

human lung muco epidermoid NCI-H292 cells

exposed to bioaerosols from a composting plant.

Different types of microorganisms were detectable



in the composting plant, using the agar culture

method. Real-time polymerase chain reaction was

used to quantify the level of Aspergillus fumigatus

and the profile of remodeling genes. The real-time

PCR results indicated that the amount of A.

fumigatus in the composting hall was less than

10(2) conidia (Alvarado-Esquivel, 2013). The

epidemiology of Helicobacter pylori (H. pylori)

infection in waste pickers had not been previously

studied. This study aims to determine the

association of H. pylori seropositivity and waste

picking activity; and to determine socio-

demographic, clinical, work, and behavioral

characteristics associated with Helicobacter

pylori seropositivity in waste pickers. Through a

case-control study design, we examined 90 waste

pickers and 90 age and gender-matched control

subjects for the presence of anti- H. pylori positive

and H. pylori negative waste pickers. Logistic

regression analysis showed that the duration

(years) in the waste picking activity was positively

associated with H. pylori exposure (OR = 2.76; 95

% CI: 1.22 - 6.25; P = 0.01). In contrast,

consumption of alcohol was negatively associated

with H. pylori exposure (OR = 0.27; 95 % CI:

0.09 - 0.78; P = 0.01). Other work or behavioral

characteristics of waste pickers including washing

hands before eating, eating from the garbage,

animal contacts, consumption of unpasteurized

milk, unwashed raw vegetables, fruits, or

untreated water, and contact with soil were not

associated with H. pylori exposure .This is the first

report on the seroprevalence of H. pylori infection

among waste pickers and the factors contributing

to such exposure. Results warrant for further

research (Kim et al., 2011).

Food waste generated in Korea is rich in

carbohydrate as high as 65 % of total solids. Using

the food waste, the feasibility of ethanol

production was investigated in a laboratory scale

fermentor. Hydrolyzing enzymes pretreatment

including carbohydrase, glucoamylase, cellulase

and protease were tested for hydrolysis of food

waste. For simultaneous saccharification and

fermentation (SSF), the ethanol yield was 0.31 g

ethanol/g total solids. During the continuous

operation of SHF, the volumetric ethanol

production rate was 1.18 g/l with an ethanol yield

of 0.3 g ethanol/g total solids. For SSF process,

the volumetric ethanol production rate was 0.8 g/h

with an ethanol yield of 0.2g ethanol/g total solids

(Li et al., 2008). With increasing attention paid to

the recycling use of aged refuse as vegetated soil,

it is important to check its possible polluting risk

and probe the tolerance of plant system to its

stress. The results implicate that aged refuse might

cause environmental stress on plant system, but

the polluting risk mainly resulted from smaller

particle-size samples. Therefore, the critical point

of utilizing aged refuse as vegetated soil focused

on analyzing its particle-size distribution, and

screening out appropriate particle-size samples

(Ma et al., 2008). Kitchen garbage was chosen to

produce ethanol through simultaneous

saccharification and fermentation (SSF) by

Zymomonas mobilis. Plackett Burman design was

employed to screen affecting parameters during

SSF process. This method could save the ethanol

production cost and benefit for the recycle of

kitchen garbage (Alvarado - Esquivel, 2013).

Municipal waste is a potential source of infection

for Toxoplasma gondii as it may contain

contaminated meat with parasite tissue cysts and

cat excrement with parasite oocysts. Therefore, we

sought to determine the prevalence of T. gondii

infection and associated characteristics in two

populations exposed to municipal solid waste in

Durango, Mexico. Ninety waste pickers and 83

waste workers of Durango City, Mexico were

examined for T (i) waste pickers may represent a

risk group for T. gondii infection; (ii) lack of

education might be a contributing factor for T.

gondii infection; (iii) the higher the exposure

to garbage, the higher the sero prevalence of T.

gondii infection; (iv) Eating food products from

the garbage may represent an important route for

T. gondii infection (Sakai et al., 2001). A

phenotypic characterization of seventeen Bacillus

strains isolated from aerobic thermo philic

bioreactors of a food waste processing company

was carried out, using fatty acid and enzymatic

activity profile. However; enzymatic activities

numerical analyses indicated that these three

Bacillus species were more homogeneous

regarding this phenotypic characteristic (Chen et

al., 2017).



Figure – 5: Feasibility of using kitchen waste as

future substrate for biofuel/bioethanol

production

4. Digestion of garbage in to gold waste

The production of solid waste has become

a common problem worldwide. Solid wastes are

the organic and inorganic waste materials formed

through various forms in the society, which is

losing their value to the users. Inappropriate

dumping of these wastes pollutes the crucial

components of the living environment like air,

land and water globally. This problem has become

more acute in developing nations like in India

compared to developed nations. Solid waste is

normally defined as unwanted solid, semi-solid

material ensuing from human or animal activities.

It is a diverse form of wastes, originating from

industrial or agricultural, household, commercial

activities. Solid waste comprises of Industrial

Waste (IW), Hazardous Waste (HW), Municipal

Solid Waste (MSW), Electronic waste (E-waste),

Bio-Medical Waste (BMW) which depend on their

supply & characteristics. On one hand, the eco-

friendly components of solid waste could be

useful as secondary source for production

processes. Some of the toxic and harmful

ingredients cause a danger if not processed

appropriately. Composting, Recycling, energy

conversion from wastes, and land filling are some

of the basic moves toward waste management

(Leslie, 2017). The renewable feedstock derived

biodegradable plastics are important in various

industries such as packaging, agricultural, paper

coating, garbage bags and biomedical implants

This review reported the degradation study of

various existing biodegradable plastics along with

the potent degrading microbes (bacteria and

fungi). Patents available on plastic biodegradation

with biotechnological significance is also

summarized in this paper This paper assesses that

new disposal technique should be adopted for the

degradation of polymers and further research is

required for the economical production of

biodegradable plastics along with their enzymatic

degradation (Mu et al., 2018).

The anaerobic digestion of single fruit and

vegetable wastes (FVW) can be easily interrupted

by rapid acido genesis and inhibition of

methanogen, and the digestion system tends to be

particularly unstable at high solid content. The

ammonium could neutralize the accumulated

volatile fatty acids (VFAs) and prevent the pH

value of the digestion system from rapidly

decreasing. Co-digestion of FVW and WAS with

TS ratio of 60:40 gave the highest biogas yield of

562 ml/g-VS and the highest methane yield of 362

ml/g-VS. Key parameters in the digestion process,

including VFAs, pH, enzyme activity & microbial

activity, were also examined (Lopez et al., 2015).

Leptospirosis is a serious infectious disease that

has linked to occupational activities such as

farmers, sewer workers, garbage collectors,

butchers and veterinarians. It is acquired through

contact with major reservoirs like rodents, pets, or

the environment contaminated by their urine. In

Paraguay, there are no published reports on the

seroprevalence of Leptospirosis related to work

activity. To determine the seroprevalence and

factors associated with transmission of

Leptospirosis in urban sanitation workers. We

recommend the improvement of prevention

strategies and health promotion in this population

(Skariyachan et al., 2015).

Industrialization and urbanization have led

to massive accumulation of plastic garbage all

over India. The persistence of plastic in soil and

aquatic environment has become ecological threat

to the metropolitan city such as Bangalore, India.

Present study investigates an eco-friendly,

efficient and cost-effective approach for plastic

waste management by the screening of novel



microbial consortia which are capable of

degrading plastic polymers. Plastic contaminated

soil and water samples were collected from six hot

spots of urban and rural areas of Bangalore. The

computational docking studies suggested that

polyethylene glycol and polystyrene present in the

plastics might have good interaction towards the

microbial lipase with stable binding and

interacting forces which probably could be one of

the reasons for the degradative mechanisms (Stone

and Morris, 2014).

The proteasome is a cellular machine

found in the cytosol, nucleus and on chromatin

that performs much of the proteolysis in

eukaryotic cells. Recent reports show it is enriched

at sites of double-stranded DNA breaks (DSBs) in

mammalian cells. What is it doing there? This

review will address three possibilities suggested

by recent reports: in degrading proteins after their

ubiquitination at and eviction from chromatin; as a

deubiquitinase, specific to the antagonism of

ubiquitin conjugates generated as part of the

signalling of a DSB; and as a functional

component of DNA repair mechanism itself.

These findings add complexity to the proteasome

as a potential therapeutic target in cancer

treatment (Daneshzadeh Tabrizi et al., 2010). It is

well known that use of low cost and abundant

waste materials in microbial fermentations can

reduce product costs. Kitchen wastes disposed of

in large amounts from cafeterias, restaurants,

dining halls, food processing plants, and

household kitchens contain high amounts of

carbohydrate components such as glucose, starch,

and cellulose Thus, it was concluded that product

cost can be lowered to a large extent if (1) kitchen

wastes are used as a substrate, (2) no fermentation

nutrient is used, and (3) hydrolysis time is applied

for about 6 hrs. Further optimization study is

needed to increase the yield to higher levels. Thus,

hypothersized that occupational exposure to

bioaerosols can increase leakage of Haws assessed

with four interview-based indicators and by

preliminary endotoxin measurements using the

Limulus amoebocyte lysate assay. No effect was

found in garbage collectors. Smoking increased

serum SP-D. No clinically relevant correlation

between spirometry results and SP-D

concentrations appeared. These results support the

hypothesis that inhalation of bioaerosols, even at

low concentrations, has a subclinical effect on the

lung-blood barrier, the permeability of which

increases without associated spiro metric changes

(Stroikin et al., 2005). The effect of various

pretreatment methods on acidogenesis of food

waste from a cafeteria was investigated. Thermal,

enzymatic, and combined thermal-enzymatic

batch pretreatment were conducted the food waste

after 60 min of thermal processing followed by 0.1

% (v/v) enzymatic treatment. Increase in VFAs

production of the fermenter was over 38 %

compared to the control fermenter without any

pretreatment. The proteasome is an abundant

multicatalytic enzyme complex present in the

cytoplasm and nucleus of all eukaryotic cells

(Daneshzadeh Tabrizi, 2010).

The primary function of the proteasome is

to degrade proteins. This article discusses the

structure and function of the proteasome, and its

role in malignant cells and as a therapeutic target

(Scott and Firth, 2004). Nature has always been

efficient at saving energy and preventing waste. A

good example of the thriftiness of nature is the

recycling of receptors that mediate the transport of

hydrolases to the lysosome in animal cells or to

the vacuole in plants and fungi. By actively

recycling these receptors, they are saved from

degradation in the "garbage can" of the cell--the

lysosome or vacuole. Until recently, this process

has been relatively poorly understood. Now,

through a fusion of yeast genetics and mammalian

cell biology, new insights have been gained into

the molecular mechanisms that underlie the

Endosome to Golgi membrane - trafficking

pathway (Savoca and Huber, 2004). The mannose

6-phosphate/insulin-like growth factor-II receptor

(M6P/IGF-IIR) is an intriguing protein with

multiple ligands and functions. Approximately 90

- 95 % of the receptor is located intracellularly,

with 5 - 10 % being on the cell surface (Garber,

2002). The proteasome, a large protease complex

in cells, is the major machinery for protein

degradation. Therefore, proteasome inhibition

represents a new and promising frontier in

immunosuppressant development Degradation of

yard wastes by Coprinus truncorum growing in a



vertical aereated bioreactor or in flasks was

studied. The measurements of pH and soluble

proteins during the incubation period were suitable

indicators of the degradation process by C.

truncorum (Wang and Cui, 2001). In order to

evaluate the suitability of cyto pathological criteria

in isolated fish hepatocytes as endpoints in

(eco)toxicological research, liver cells isolated

from rainbow trout (Oncorhynchus mykiss) by

collagenase perfusion were exposed in vitro for up

to 5 days to sublethal dilutions of two seepage

water samples collected from garbage dumps. (3)

Prolonged exposure of isolated hepatocytes in

combination with ultrastructural and biochemical

investigations as a sensitive tool to detect adverse

effects at environmentally relevant toxicant

concentrations (Zahn et al., 1995).

5. Organic fruit waste as soil health and

supplement

In wastewater treatment, due to a lack of

complex digestive stems, bacteria need to pre-

digest the potential food sources such as organic

and inorganic materials in wastewater outside their

cell boundaries first. To accomplish this pre-

digestion bacteria excrete enzymes through their

enveloping membrane with its supportive cell wall

into the surrounding environment These “extra –

cellular enzymes” are reasonably stable, highly

resistant to chemicals, and are able to function

over a relatively broad temperature range, in order

to survive in then environment outside the

protection of the cell‟s wall (Pande and Nagernaik,

2011). Additives in wastewater treatment are

available as biological and chemical additives.

Chemical additives may be harmful to the

environment and are generally discouraged

banned because of strong acids, bases or toxic

contents, and possibly result in adverse effects on

system components, the soil structure, or ground

water quality. Biological additives have significant

beneficial impacts and do not directly harm

traditional onsite systems. Example of types of

additives used as flocculants are organic polymer,

aluminum salts, lignite coke, loam – sand mixture,

coal, bentonite, limeston ,chemical polymer, and

poly electrolytes (Veeken and Hamelers, 1999).

The garbage enzymes produced from

preconsumer organic waste containing multi

hydrolytic enzyme activity which helps to

solubilize the waste activated sludge. The

continuous production of garbage enzyme and its

scaling up process need a globe optimized

condition. Garbage enzyme produced from 6 g

pineapple peels: 4g citrus peels pre-treated with

ultrasound for 20 minutes had shows the higher

hydrolytic enzymes activity. These optimized

conditions can be successfully used for large scale

production of garbage enzyme with the higher

hydrolytic enzyme activity. There are many

problems such as low soil organic matter,

available nutrients and microbial activity,

compaction, and poor tillage properties for a

newly reclaimed cultivated land, and the

establishment of a fast, effective measure for

improving soil fertility quality is of importance to

enhance the quality and production performance

of the newly cultivated land. A field experiment

was carried out to observe the effect of organic

wastes on soil fertility of a newly reclaimed

cultivated land, and compared the differences of

different types of urban organic wastes. The field

experiment included nine treatments, i.e., pig

manure, chicken manure, rice straw, vegetable

harvest residue, urban sludge, biogas residue,

manure + rice straw compost, garbage compost

and control without organic fertilizer at annual

application rate of 30 t.hm-2

, and ran for three

consecutive years. Overall, effects of organic

wastes on soil fertility decreased in the order of

pig manure + rice straw compost > chicken

manure > pig manure > biogas residue >

garbage compost > rice straw > urban sludge >

vegetable harvest residue, and the effects on soil

pollution increased in the sequence of rice straw <

vegetable harvest residue < pig manure + rice

straw compost < biogas residue < chicken manure

< pig manure < garbage compost < urban sludge

(Xu et al., 2016; Arun and Sivashanmugam,

2017).

The amount of food waste generated from

modern societies is increasing, which has imposed

a tremendous pressure on its treatment and

disposal. It can be concluded that appropriate

portions of different types of food waste could



satisfy basic nutritional requirements of lower

tropic level fish species such as grass carp and

tilapia. Upgrading the fish pellets by adding

different supplements mentioned above could

further elevated the quality of feeds, leading to

higher growth rates, and enhanced immunity of

fish. Health risk assessments based on the major

environmental contaminants (mercury, PAHs and

DDTs) in fish flesh showed that fish fed food

waste based pellets are safer for consumption,

when compared with those fed commercial feed

pellets (Wong et al., 2016). The Wangyang River

(WYR) basin is a typical wastewater irrigation

area in Hebei Province, North China. This study

investigated the concentration and distribution of

six priority phthalate esters (PAEs) in the

agricultural soils in this area. Among the DEHP

concentrations, the highest DEHP concentration

was found at the sites close to the villages; this

result suggested that dense anthropogenic

activities and random garbage disposal in the rural

area are possible sources of PAEs. The PAE

concentrations were weakly and positively

correlated with soil organic carbon and

soil enzyme activities; thus, these factors can

affect the distribution of PAEs. This study further

showed that only dimethyl phthalate (DMP)

concentrations exceeded the recommended

allowable concentrations; no remediation

measures are necessary to control the PAEs in the

WYR area. However, the PAEs in the topsoil may

pose a potential risk to the ecosystem and human

health in this area. Therefore, the exacerbating

PAE pollution should be addressed (Zhang et al.,

2015). Short-chain fatty acids (SCFAs) are the

valuable products derived from the anaerobic

fermentation of organic solid waste. The activities

of key enzymes such as hydrolytic and acid-

forming enzymes were greatly promoted due to

the presence of APG. These results demonstrated

that the enhanced mechanism of SCFAs

production should be attributed to the acceleration

of solubilization and hydrolysis, enhancement of

acidification and inhibition of methanogenesis by

APG (Zhao et al., 2015).

Infection with Leptospira may occur by

contact with Leptospira - infected animals. Waste

pickers are in contact with rodents and dogs while

picking in the garbage. Whether waste pickers are

at risk for Leptospira infection is largely unknown.

This study was aimed to determine the association

of Leptospira sp. IgG seroprevalence with the

occupation of waste picking, and to determine the

epidemiological characteristics of the waste

pickers with Leptospira exposure. Through a case-

control study, we determined the seroprevalence

of anti-Leptospira IgG antibodies in 90 waste

pickers and 90 age- and gender-matched control

subjects in Durango City, Mexico using

an enzyme immunoassay. This is the first study

about Leptospira exposure in waste pickers.

Results suggest that waste pickers are not at

increasing risk for Leptospira exposure in

Durango City, Mexico. Further research with a

larger sample size to elucidate the association of

Leptospira exposure with waste picking activity is

needed (Alvarado-Esquivel et al., 2015). We

investigated L-lactic acid production in static

batch fermentation of kitchen refuse using a

bacterial consortium from marine-animal-resource

(MAR) composts at temperatures ranging from 30

to 65 °C. At relatively low temperatures butyric

acid accumulated, whereas at higher temperatures

L-lactic acid was produced. This is the first report

of the achievement of 100 % optical purity of L-

lactic acid using a bacterial consortium (Tashiro et

al., 2013). Pro biotic are live microorganisms that

exert beneficial effects on their host. A high

survival rate during gastrointestinal transit and

storage is often desirable The viability of

Lactobacillus acidophilus FTDC 1331

incorporated in carriers containing agro wastes

was more stable during storage at 4 °C for 28 days

compared with the control (P < 0.05) (Lew et al.,

2001).

Kitchen waste from dining rooms accounts

for a considerable proportion of municipal

solid garbage, and economical recycle ways are

needed to be developed. This study investigated

gluco amylase production from kitchen waste and

the feasibility of kitchen waste hydrolysis by the

crude enzymes produced.



Fig – 6: Management of waste generation

derived through plant and animal food supply

The optimum enzyme dose 8 %

(crude enzyme/kitchen waste, w/w) was not too

big, and was sufficient to hydrolyse 10 % (dry

basis) sKW slurry to produce a maximum amount

of reducing sugar of 55.4 g L-1

(Wang et al.,

2001). A stable mutant flocculent yeast strain of

Saccharomyces cerevisiae KRM-1 was isolated

during repeated-batch ethanol fermentation using

kitchen refuse as the medium. Cell recycle by

flocculation was fast and convenient, and could

therefore be applicable for industrial-scale ethanol

productive (Ma et al., 2009). Uncontrolled

combustion due to garbage recycling is a

widespread activity among slum dwellers in

distressed economy countries and has been

indicated as a major source of dioxin

contamination. It was concluded that the ELISA

method performed very well as a screening tool to

prioritize samples for instrumental analysis, which

allows cutting down costs significantly (Trindade

et al., 2008). Aging (senescence) is characterized

by a progressive accumulation of macromolecular

damage, supposedly due to a continuous minor

oxidative stress associated with mitochondrial

respiration. Aging mainly affects long-lived post

mitotic cells, such as neurons and cardiac

myocytes, which neither divide and dilute

damaged structures, nor are replaced by newly

differentiated cells (Silva et al., 2006). Cellular

manifestations of aging are most pronounced in

postmitotic cells, such as neurons and cardiac

myocytes. This interrelated mitochondrial and

lysosomal damage irreversibly leads to functional

decay and death of postmitotic cells (Diorio et al.,

2001).

6. Miscellaneous

For the past millennium, the world has run

on crude oil and coal as the main energy source. In

the past decade, the price of crude oil has doubled,

and with climate change imminent, the world has

to re-evaluate its economic growth and energy

policies. South Africa, as part of Africa, has the

added burdens of rising unemployment and

poverty and the need to decouple its economy

from fossil fuels. The demand to replace

conventional industrial processes with those that

generate fewer or no pollutants is increasing as a

result of the need to minimize anthropogenic

environmental impacts (Demarche et al., 2012). In

January 2014, the South African Department of

Science and Technology revealed the national

Bio-Economy Strategy 5, in this document, the

term bio-economy „encompasses biotechnological

activities and processes that translate into

economic outputs, particularly those with

industrial application‟. The vision for South Africa

sees the bio-economy contributing significantly to

the country‟s gross domestic product by 2030

through the creation and „growth of novel

industries that generate an develop bio-based

services, products and innovations‟5. The

potential of a thriving bio-economy will affect the

country on a macro-economic scale, making South

Africa internationally competitive (especially in

the industrial and agricultural sectors) by creating

more sustainable jobs, linking the countries first

and second economies, enhancing food security

and creating a greener economy. The strategy

presents a framework for the development of a

thriving bio-economy, in which collaboration

between role players (including, among others, the

biotechnology sector as a whole, environmental

agencies and social scientists) is the key to

success. The three key economic sectors identified

for inclusion in the strategy are agriculture, health

and industry (Department of Science and

Technology, 2013).

Fruit processing (canning, juicing,

winemaking and drying) generates large quantities

of waste, both solid and liquid. For example,



approximately 25 - 35 % of processed apples (dry

mass), 50 % of citrus and 20 % of grapes end up

as waste. The solid waste, often called pomace, is

the portion of the fruit that is not utilized, such as

skins, pips and fibres. The pomace has high lingo

cellulose content and is very recalcitrant to

degradation. For example, wastewater with a

chemical oxygen demand (COD) of less than 400

mg/L can be used for irrigation at volumes of up

to 500 m3, while irrigation volumes may not

exceed 50 m3 on any given day if the COD is

between 400 mg/L and 5000 mg/L. The average

COD of wastewater in the juicing and canning

industries is often as high as 10000 mg/L and

therefore requires extensive treatment before

discharge into the environment (Al – Jasser,

2009).

7. Conclusion Enzymes are being known to mankind

since the ancient human civilization. The use of

enzymes had been done intensively in different

fields especially, in ancient brewing and other

uses. But since the 18th

century it has been

technically known to us as enzymes. Many

scientists had tried to study the use of enzymes,

and from their pioneer work, we have come to

know about its power and utility in our daily life.

Today different types of enzymes are being

manufactured by many big companies and being

sold for their important role in different industries

like food, dairy, detergent, and chemical as well as

for their important lifesaving therapeutically

application. Different microbes as well as other

model systems are extensively used for the

production of these important enzyme bio-

cleaning agents. Since then many microorganisms

and their enzymes with unique function have also

been discovered by means of extensive screening,

and now they are commonly used in different

fields. Development of these biocleaning enzymes

has been at least as extensive as those for

industrial applications thus reflecting the

magnitude of the potential rewards of this sector in

the near future. This review especially emphasizes

the production technologies and important wide

spectrum role of garbage based enzymes in

various sectors of industries. There is an indeed

need of future research in these biomolecules

which will later be beneficial for the mankind in

their relevance. Similarly, further research and

clear demonstrations for applications of human-

friendly, environmentally-sustainable techniques

and technologies for the conservation of

biodiversity and environment. The confirmation of

the safety of these technologies will also lead to

the standardization of protocols, evaluation of

end-user costs, and commercialization and,

therefore, the production development of

innovative “ready to use” products without

affecting our earth and environment.

Perspectives

The enzyme bio-cleaning products that are

easily-applied, ready-to-use, and include fast

application, will finally become a reality with

scientific approval. At that time appropriate cost-

to-benefit evaluation will confirm enzyme bio-

cleaning technology to not only be

environmentally sustainable, but also

economically viable. We believe that on the basis

of the before mentioned considerations, more

Indian Community projects should finance

research on these matters.

Author Contributions

Each author focused on a topic, performed

an accurate search of papers, and wrote the

relevant section.

Acknowledgement

We take this opportunity to acknowledge

sincere thanks to our respected chairman, Dr S.C.

Kulshreshtha, Honorable Executive Director Dr

B.K Tyagi, Director Dr. R.S. Saxena, Shri Ram

Group of Colleges Muzaffarnagar, U.P. India for

providing necessary facility and tools to carry out

the research dissertation work for post graduate

students of M.Sc Biotechnology.

Source of support – Nil.

Conflict of interest – None declared.

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DOI Number DOI: 10.22192/lsa.2018.4.2.1

How to Cite this Article:

Ashwani Kumar, Vaishali Dhiman, Anuradha Saini and Amit Panwar. 2018. An exculsive

review on production, analysis and application of Enzyme Bio-cleaners from fruits and

vegetable wastes. Life Science Archives, 4(2): 1292 – 1308.

DOI: 10.22192/lsa.2018.4.2.1

http://www.jpsscientificpublications.com/

life science archives (lsa)by dr. rosukon can be classify as a complex organic substance of protein...

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