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Study on Tannase Produced by Aspergillus Niger and Propyl Gallate Biosynthesised by Immobilized Aspergillus Niger Cells in Organic Phase

Author: ZhouJiDong
Tutor: LiYongQuan
School: Zhejiang University
Course: Genetics
Keywords: Aspergillus niger The organic phase Alginate Immobilized Aspergillus niger cells Tannase Compound Mutation Biosynthesis Propyl gallate
CLC: TQ920
Type: Master's thesis
Year: 2002
Downloads: 332
Quote: 10
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Abstract


In this study, producing Aspergillus niger strains tannin as the starting bacteria, enzyme production through mutation breeding and fermentation process optimization to improve the ability of enzyme production strains, and then be immobilized cells, used in the biosynthesis of the organic phase antioxidant gallic gallate (propyl gallate, PG) studies. The organic phase immobilized cell PG biosynthesis research at home and abroad is the first time, this study in the academic and practical applications are of great significance. In previous studies, based on the constructed tannin enzyme activity assay methods: PG as a substrate for the determination of 270nm wavelength ultraviolet region by propyl gallate concentration and absorbance values ??calculated from the linear relationship between the bottom The reduction was thus measured enzyme activity tannin. This method is simple, rapid, sensitive and reliable, reproducible, high reliability, suitable for intracellular tannase enzyme activity determination. Microwave mutagenesis and chemical mutagenesis through a combination of composite breeding, ultimately yielding Aspergillus niger tannase L 21 strains, enzyme production up 130.6U/ml. In the fermentation of the strain under conditions of enzyme production process parameters including the concentration of tannic acid, carbon, nitrogen, carbon and nitrogen ratio, initial pH, temperature and other factors speed and culture studies, and then use L 27 (3 13 ) orthogonal to the above seven major factors to be optimized. The optimum fermentation conditions, Aspergillus niger L 21 strains obtained tannase enzyme 225.8U/ml. Through six kinds of Aspergillus niger cell immobilization methods comprehensive comparison, the final selection of calcium alginate embedding. Given immobilized cells in mycelial growth and enzyme production aspects are exhibited hysteresis effect, so the Aspergillus niger L 21 immobilized cell fermentation conditions for enzyme production in concentration of sodium alginate, CaCl 2 < / sub> concentration of spore concentration gel, plastic beads inoculation, tannic acid concentration and volume of liquid that six factors were studied, and the use of L 18 (3 7 ) orthogonal test optimization. After optimization, Aspergillus niger L 21 immobilized cells and free cells not only has a similar enzyme production process and the pre-peak enzyme production, but also exhibit a balance over a longer period of free cells. Finally, the immobilized Aspergillus niger L 21 cell biocatalysis in organic synthesis PG. By the choice of the reaction system and Preliminary Optimization of synthesis parameters, obtained about 20% yield of PG.

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