Applied and Industrial Applied and Industrial Microbiology Microbiology

(BISC&BTEC6343)(BISC&BTEC6343)

BIOTECHNOLOGY IS UTILIZATION OF BIOTECHNOLOGY IS UTILIZATION OF BIOLOGICAL SYSTEMSBIOLOGICAL SYSTEMS

OR PART OF BIOLOGICAL SYSTEMS OR PART OF BIOLOGICAL SYSTEMS TO GET ANY PRODUCT OR SERVICETO GET ANY PRODUCT OR SERVICE

Employing plant cells, animal cells and Employing plant cells, animal cells and microorganisms not only to microorganisms not only to manufacture goods and medicines manufacture goods and medicines that are useful to mankind, that are useful to mankind, but alsobut also to to generate products and procedures generate products and procedures which will improve the quality and which will improve the quality and health of livestock, agricultural crops health of livestock, agricultural crops and our environment at large.and our environment at large.

Chemistry

Biology Engineering

Ch.Eng.

Bioeng.

Biochemistry

Biotech.

Biotechnology involves integration Biotechnology involves integration of such advanced disciplines asof such advanced disciplines as::

History of BiotechnologyHistory of Biotechnology

Dates back to ancient times. Alcoholic beverages, Dates back to ancient times. Alcoholic beverages, bread, cheese are first examples of unconscious use bread, cheese are first examples of unconscious use of biotechnology.of biotechnology.

Visualization of cells by Leeuwenhoek in 17th Visualization of cells by Leeuwenhoek in 17th century. The discovery of role of yeast in alcoholic century. The discovery of role of yeast in alcoholic fermentation by Pasteur in mid 1800s, utilization of fermentation by Pasteur in mid 1800s, utilization of pure yeast cultures in brewery by Hansen, detection pure yeast cultures in brewery by Hansen, detection of fermentation enzymes in yeast by Buchner in late of fermentation enzymes in yeast by Buchner in late 1800s. Then, early 20th century saw the first 1800s. Then, early 20th century saw the first biological sewage treatment plants. biological sewage treatment plants.

Discovery of penicillin by Flemming triggered Discovery of penicillin by Flemming triggered the birth of modern biotechnology. This the birth of modern biotechnology. This introduced the requirements for aseptic introduced the requirements for aseptic fermentation (bioprocess) techniques where fermentation (bioprocess) techniques where contaminating microbes are excluded. Other contaminating microbes are excluded. Other antibiotics, vaccines, single cell proteins, antibiotics, vaccines, single cell proteins, vitamins, nucleotides have been produced vitamins, nucleotides have been produced commercially ever since, with more commercially ever since, with more sophisticated fermentation and extraction sophisticated fermentation and extraction methods. methods.

In recent years, molecular biology techniques In recent years, molecular biology techniques and Genetic engineering have made possible; and Genetic engineering have made possible; the gene manipulations for production of highly the gene manipulations for production of highly productive strains, the utilization of bacteria for productive strains, the utilization of bacteria for production of rare chemicals like insulin, production of rare chemicals like insulin, interferon, growth hormone, viral antigen etc. interferon, growth hormone, viral antigen etc. The discovery of hybridoma technology The discovery of hybridoma technology (production of monoclonal antibodies) in 1975. (production of monoclonal antibodies) in 1975.

Another developing area is bioprocess Another developing area is bioprocess engineering, investigating the ways to optimize engineering, investigating the ways to optimize fermentation process and products in the field of fermentation process and products in the field of biotechnology. biotechnology.

Biotechnologically Biotechnologically important important

microorganismsmicroorganisms

BacteriaBacteria         belong to kingdom Monera belong to kingdom Monera         are procaryotes, genetic material is not surrounded are procaryotes, genetic material is not surrounded

by a special nuclear membrane by a special nuclear membrane         single celled organisms single celled organisms         their shape may be coccus, bacillus, spiral, their shape may be coccus, bacillus, spiral,

square, star shaped and individual bacteria may form square, star shaped and individual bacteria may form chains, clusters, pair etc.chains, clusters, pair etc.

generally reproduce by binary fission         generally reproduce by binary fission                 some bacteria use organic compounds, some use some bacteria use organic compounds, some use

inorganic compounds for nutrition, others make inorganic compounds for nutrition, others make photosynthesis to manufacture their own food photosynthesis to manufacture their own food

        important tool for recombinant DNA technology important tool for recombinant DNA technology

A typical bacterial cell is shown below.

Scanning electron micrograph of E.coli

        FUNGIFUNGI belong to kingdom Fungi belong to kingdom Fungi         are eucaryotes, genetic material surrounded by are eucaryotes, genetic material surrounded by

nuclear membrane nuclear membrane         include unicellular yeast, multicellular molds and include unicellular yeast, multicellular molds and

large multicellular mushrooms large multicellular mushrooms         reproduce sexually or asexually reproduce sexually or asexually         absorb dissolved solutions of organic compounds absorb dissolved solutions of organic compounds

from environment from environment         good source for enzymes, polysaccharides, lipids good source for enzymes, polysaccharides, lipids

or other valuable bioproducts. or other valuable bioproducts.         used to supply human and animal diet with its high used to supply human and animal diet with its high

protein content and vitamins (single cell proteins, protein content and vitamins (single cell proteins, SCP) SCP)

        ProtozoaProtozoa belong to kingdom Protista belong to kingdom Protista         are eucaryotes are eucaryotes         unicellular unicellular         movement through pseudopods, flagella or movement through pseudopods, flagella or

cilia cilia         reproduce sexually or asexually reproduce sexually or asexually         feed upon bacteria and small particulate feed upon bacteria and small particulate

nutrients, some are part of normal microbiota nutrients, some are part of normal microbiota of animals of animals

AlgaeAlgae

some belong to kingdom Protista, some some belong to kingdom Protista, some belong to Plantae belong to Plantae

        are eucaryotes are eucaryotes         reproduce sexually or asexually reproduce sexually or asexually         photoautotrophs; they use light as source of photoautotrophs; they use light as source of

energy. energy.         good source for enzymes, pigments, lipids good source for enzymes, pigments, lipids

etc. etc.

VirusesViruses

        have DNA or RNA as genetic material have DNA or RNA as genetic material         obligatory intracellular parasites, they obligatory intracellular parasites, they

multiply inside living cells. multiply inside living cells.         for multiplication, they invade host cell and for multiplication, they invade host cell and

by directing its metabolic machinery by directing its metabolic machinery synthesize viral nucleic acids and enzymes. synthesize viral nucleic acids and enzymes.

        are used for vaccine production and serve are used for vaccine production and serve as vectors for gene transfer as vectors for gene transfer

Industrial chemicals

chemicals Microbial sources

ethanol Saccharomyces

acetic acid Acetobacter

glycerol Acetobacter

isopropanol Clostridium

acetone Clostridium

Enzymes

Enzymes microbial sources

alpha amylase

Bacillus licheniformis

Cellulases Phanerochaete chrysosporium

Lipases Aspergillus niger

Some examples of substances synthesized by microorganisms

Amino acids

amino acids Microbial sources

L-Arginine Brevibacterium flavum

L-Leucine Brevibacterium lactofermentum

L-Phenylalanine

Brevibacterium lactofermentum

L-Serine Corynebacterium hydrocarboclastus

L-Tryptophan Brevibacterium flavum

Antibiotics

Antibiotics microbial sources

Cephalosporin C Cephalosporium acremonium

Chloramphenicol Streptomyces venezuelae

Penicillin Penicllium chrysogenum

Tetracycline Streptomyces aureofaciens

Pharmacologically important compounds

compounds Microbial source

Dopastin Pseudomonas sp.

Esterastin Streptomyces lavendulae

Naematolin Naematoloma fasciculare

Phialocin Phialocephala repens

Polymers

polymers microbial source

Cellulose Acetobacter sp.

Dextran Acetobacter sp.

Xanthan Xanthomanas campestris

Food and Beverages

Products microbial source

Bread Saccharomyces cerevisiae

beer and wine Saccharomyces cerevisiae

soy souce Aspergillus oryzae

Yoghurt Lactobacillus and Streptococcus

Single cell proteins

product (biomass) microbial source substrate used

Saccharomycodes lipolytica

Saccharomycodes lipolytica n-alkenes

Fusarium graminearum

Fusarium graminearum polysaccharides

Pruteen Methylophilus methylotrophus

methanol

algae Algae from CO2 /sunlight

Microbial products can be classified Microbial products can be classified asas

The microbial cells themselves; biomass The microbial cells themselves; biomass production production

Primary metabolites; which are produced by Primary metabolites; which are produced by cells to live and grow. Alcohol, amino acids, cells to live and grow. Alcohol, amino acids, nucleotides, fats, vitamins and enzymes are nucleotides, fats, vitamins and enzymes are examples of commercially important products examples of commercially important products

Secondary metabolites; are not necessarily Secondary metabolites; are not necessarily required for life. Antibiotics and various required for life. Antibiotics and various alkaloids are the most important secondary alkaloids are the most important secondary metabolites metabolites

Advantages of microbial Advantages of microbial systemssystems

Growth is fastGrowth is fast Mass cultivation is possible in large Mass cultivation is possible in large

fermentersfermenters Medium can be designed with cheaper Medium can be designed with cheaper

components.components. Genetic manipulations are widely and easily Genetic manipulations are widely and easily

performed. performed.

CHARACTERISTICS IMPORTANT IN CHARACTERISTICS IMPORTANT IN MICROBES USED IN INDUSTRIAL MICROBES USED IN INDUSTRIAL

MICROBIOLOGY AND MICROBIOLOGY AND BIOTECHNOLGYBIOTECHNOLGY

i.i. The organism must be able to grow in a The organism must be able to grow in a simple medium and should preferably not simple medium and should preferably not require growth factors. require growth factors.

ii.ii. The organism should be able to grow The organism should be able to grow vigorously and rapidly in the medium in use. vigorously and rapidly in the medium in use.

iii.iii. It should also produce the desired It should also produce the desired materials, whether they be cells or materials, whether they be cells or metabolic products, in as short a time as metabolic products, in as short a time as possible.possible.

iv.iv. Its end products should not include toxic Its end products should not include toxic and other undesirable materials, especially and other undesirable materials, especially if these end products are for internal if these end products are for internal consumption.consumption.

v. The organism should have a reasonable genetic, v. The organism should have a reasonable genetic, and hence physiological stability.and hence physiological stability.

vi. The organism should lend itself to a suitable vi. The organism should lend itself to a suitable method of product harvest at the end of the method of product harvest at the end of the fermentation. fermentation.

vii. Wherever possible, organisms which have vii. Wherever possible, organisms which have physiological requirements which protect them physiological requirements which protect them against competition from contaminants should be against competition from contaminants should be used. An organism with optimum productivity at used. An organism with optimum productivity at high temperatures, low pH values or which is able high temperatures, low pH values or which is able to elaborate agents inhibitory to competitors has to elaborate agents inhibitory to competitors has a decided advantage over others. Thus a a decided advantage over others. Thus a thermophilic efficient producer would be preferred thermophilic efficient producer would be preferred to a mesophilic one. to a mesophilic one.

viii. The organism should be reasonably viii. The organism should be reasonably resistant to predators such as resistant to predators such as Bdellovibrio Bdellovibrio spp or bacteriophages. spp or bacteriophages.

ix. Where practicable the organism should ix. Where practicable the organism should not be too highly demanding of oxygen as not be too highly demanding of oxygen as aeration contributes about 20% of the cost aeration contributes about 20% of the cost of the finished product.of the finished product.

x. Lastly, the organism should be fairly easily x. Lastly, the organism should be fairly easily amenable to genetic manipulation to amenable to genetic manipulation to enable the establishment of strains with enable the establishment of strains with more acceptable properties.more acceptable properties.