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Published on: July 26, 2024
[Production of gentiooligosaccharide by recombinant beta-glucosidase]
Lingling Liu1, Song Zhu, Ting Zhu
1State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China. linglingliuwuxi@yahoo.com.cn
Objective:
Beta-glucosidase can be used to prepare gentiooligosaccharide from glucose. The purpose of this study is to obtain beta-glucosidase through DNA recombinant technology as well as to optimize the production of gentiooligosaccharide by the recombinant beta-glucosidase.
Methods:
We cloned bgl, the gene encoding beta-glucosidase from Aspergillus niger (CMI CC 324626) into the expression vector pPIC9K to construct the recombinant plasmid pPIC9K-bgl. The vector was then transformed into Pichia pastoris KM71 for extracellular overproduction of beta-glucosidase. The activity of the expressed enzyme was measured by the assay of transglucosidation reaction and the transglucosidation product was identified by HPLC and LC-MS. Furthermore, the condition for prepare gentiooligosaccharide by this recombinant beta-glucosidase is optimized.
Results:
A. niger beta-glucosidase was successfully expressed in P. pastoris and the recombinant produced gentiooligosaccharide from glucose. In addition, the main operation parameters of this enzymatic conversion were optimized. At 80% glucose, 60 degrees C, pH 4.5, 1 mmol/L K+, 60 U beta-glucosidase per gram substrate, and 48 h reaction time, the gentiooligosacharide produced reached 50 g/L.
Conclusion:
This is the first report of producing gentiooligosaccharide by recombinant beta-glucosidase.
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