谷氨酸棒状杆菌全细胞催化合成Levan果聚糖条件优化分析

doi:10.16431/j.cnki.1671-7236.2020.11.008

Abstract

Abstract: This experiment was aimed to explore a new way for microbial synthesis of Levan and optimize its synthesis conditions.Levan synthesis by whole-cell catalysis using recombinant Corynebacterium glutamicum that transferred into levansucrase,and the response surface method were used to optimize the synthesis conditions with Levan yield as the response value.Firstly,the Plackett-Burman design was used to screen out three main factors affecting yield (sucrose concentration,final concentration of ethanol in Levan extraction and Ca²⁺ concentration) from 7 related factors (sucrose concentration,bacterial concentration,transformation time,transformation pH,Ca²⁺ and Mg²⁺ concentration in PBS,and final concentration of ethanol in Levan fructan extraction).Then the three most important factors selected were used for the steepest climbing test to approach the region of maximum response value,and finally use the BBD response surface method to determine the interaction between the three main factors and the best synthesis conditions.The results showed that the average yield of Levan was 0.069 g/mL in the initial experiment.The response surface method was used to optimize the whole-cell biocatalysis of Levan by Corynebacterium glutamate.The most influential factor was sucrose concentration,the second was the final concentration of ethanol,and the last was the Ca²⁺ concentration in the transformation solution.When the sucrose concentration was 52.71%,the final ethanol concentration was 85.31%,and the Ca²⁺ concentration was 0.04 g/L,the average yield of Levan synthesized by recombination Corynebacterium glutamate was 0.238 g/mL,which was close to the model's predicted maximum value of 0.233 g/mL,and the conversion rate was about 44%.The conversion rate was 1.9 times higher than the initial test,and the yield was 3.4 times higher than the initial test.The results provided the basis for Levan industrial production,and laid the foundation for its research in animal medicine and nutrition.

Key words: Corynebacterium glutamicum; levansucrase; Levan; whole-cell biocatalysis; response surface methodology

CLC Number:

S852.23

WU Xinyu, YAN Lin, WU Hong. Optimizing Conditions Analysis of Levan by Whole-cell Biocatalysis with Corynebacterium glutamicum[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(11): 3474-3483.

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Optimizing Conditions Analysis of Levan by Whole-cell Biocatalysis with Corynebacterium glutamicum

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