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S448 Abstracts / Journal of Biotec

V4-P-120

Production of composite biomaterials for medical applicationon the basis of chitosan from Blakeslea trispora industrial waste

O.V. Kalinkevich 1,∗, A.M. Sklyar 2, S.N. Danilchenko 1, V.I. Kindya 3,A.N. Kalinkevich 1, L.F. Sukhodub 1

1 Institute of Applied Physics, NAS of Ukraine, Sumy 40030, Ukraine2 Sumy State Pedagogical University, Sumy, Ukraine3 Sumy National Agrarian University, Sumy, Ukraine

E-mail address: bio20001@yandex.ru (O.V. Kalinkevich).

The filamentous fungus Blakeslea trispora is a promising producer oflipophilic biocorrectors and adaptogens. Fungal mycelial wastes canalso be alternative sources of chitin/chitosan materials, in additionto the traditional industrial source, the crustacean waste. Moreover,the fungal chitosans have unique properties in comparison withthose derived from Crustacea. We have investigated the possibilityof production of chitosan from B. trispora mycelial wastes, comingfrom carotene production in a biotechnological plant in Ukraine.Chitosan was extracted from B. trispora using alkali extractionand characterized by physicochemical methods (FTIR spectroscopy,SEM). On the basis of the obtained fungal chitosan, compositecalcium-phosphate materials have been synthesized. These mate-rials can be used in bone defect replacement. The technologicalprocess of carotene preparation production from B. trispora biomassconsists from the following stages: (1) Preparation and growing ofinoculum. (2) Fermentation, or the process of carotene biosynthe-sis. The submerged cultivation method of producer in the fermenteris used for the carotene biosynthesis. (3) Vacuum drying of the fun-gal mycelium. (4) Extraction of carotene with oil. Residual lipidswere extracted from the industrial mycelium waste with chloro-form and methanol. Non-lipid mycelium wastes were washed with3% (w/v) KOH ca. for 30 min, then were washed 30 min with 1%(w/v) HCl. Insoluble material was treated with 60% KOH at 130 ◦C

for 3 h. Presumable chitosan was washed with distilled water toobtain neutral pH, and acetone, and then dried. The preparation ofcomposite was performed by co-precipitation method. This methodincludes the production of a dilute solution of chitosan (0.2 wt%) in1 vol% acetic acid, followed by the addition of aqueous solutions ofcalcium chloride and sodium dihydrogen phosphate. A dilute solu-tion of sodium hydroxide was slowly added to the stirred solution,bringing the pH to the value above 11. Then the suspension wasstirred for 24 h. A solid ceramic-like composite was obtained bydrying the composite hydrogel at room temperature. The obtainedcomposite material has the acceptable mechanical properties. Thechitosan/calcium phosphate composite materials obtained by thesimilar procedure and characterized by XRD had been shown tocontain the nanocrystalline hydroxyapatite as the mineral part,which is also important for the biocompatibility of the composites.We hope that fungal chitosans could be used for the production ofbioactive composite materials for bone defect replacement due totheir wound healing and haemostatic properties.

doi:10.1016/j.jbiotec.2008.07.1039

gy 136S (2008) S402–S459

V4-P-121

Analysis of the “NADH pathway” in whole cells of Enterobacteraerogenes for H2 production and its new application in NAD+

regeneration

Chong Zhang, Kun Ma, Si-Jin Li, Xin-Hui Xing ∗

Department of Chemical Engineering, Tsinghua University, Beijing100084, PR China

E-mail address: xhxing@tsinghua.edu.cn (X.-H. Xing).

The mechanism of the “NADH pathway” for H2 production in wholecells of Enterobacter aerogenes was studied. Externally added NADHwas proved to be oxidized by E. aerogenes to form 1 mol of H2and NAD+, and the formed NAD+ was then utilized by the cellsto produce H2. NADH was transformed to NAD+ by E. aerogenescells, and at the same time, the intracellular protons were reduced.Under anaerobic conditions, the reduced protons was seemed tobe supplied to a membrane bound, putatively NADH-linked [NiFe]hydrogenase. This hydrogenase was responsible for the forma-tion of H2 via the NADH pathway. While under aerobic condition,the reduced protons were reacted directly with oxygen to formH2O via the electron transport chains in the cells. By coupling theNADH oxidization in whole cells with an NAD+-dependent ADH, anew NAD+ regeneration system was constructed, reaching a totalturnover number (TTN) of 580 for the oxidization of ethanol toaldehyde.

Keywords: Enterobacter aerogenes; Hydrogenase; H2 production;NADH; NAD+; NAD+ regeneration

doi:10.1016/j.jbiotec.2008.07.1040

No. V4-P-124

Studies on media optimization for enhanced phb production &kinetics of phb granule formation in alcaligenes sp.ncim 5085using transmission electron microscopy

Abhishek Dutt Tripathi ∗, S.K. Srivastava

School of Biochemical Engineering, Institute of Technology, Banaras

Hindu University, VARANASI-221005, U.P., India

E-mail address: abhishekpam007@gmail.com (A.D. Tripathi).

Poly-3-hydroxybutyrate (PHB) is a natural biodegradable polymer(Anderson and Dawes, 1990) accumulated as an energy reservematerial by a large number of bacteria under nitrogen, phos-phorus, magnesium limited condition or in excess of carbonsource.Alcaligenes eutrophus is considered as a potent PHB pro-ducing microbe worldwide. The major problem associated withthe industrial production PHB is their high cost of production. Inthe present study, efforts were made to optimize the growth ofAlcaligenes sp.NCIM 5085 in the presence of nutrients which couldnot only decrease the production cost of PHB but also increasethe productivity. Quantitative estimation of PHB was done by Gaschromatography using NUKON gas chromatograph (Braunegg et al.,1978). TEM (Transmission Electron Microscopy) was performed tostudy the process of granule formation in microbe under nitrogenlimited condition. At the center of the cells dark stained mediationelements were seen which were considered to be nucleation site forgranule formation. TEM image also revealed the increase in cell sizeand volume to two and three folds during granule formation under

∗ Corresponding author. Tel.: +86 10 6279 4771; fax: +86 10 6277 0304.