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Extracellular expression and characterization of an α-glucosidase from Oryza sativa and its transglycosylation for synthesis of 2-O-α-D-glucopyranosyl-L-ascorbic acid
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  • Xuelian Qi,
  • Junlan Shao,
  • Yinchu Cheng,
  • Xiaoying He,
  • Yan Li,
  • Honghua Jia,
  • Ming Yan
Xuelian Qi
Nanjing Tech University
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Junlan Shao
Nanjing Tech University
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Yinchu Cheng
Nanjing Tech University
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Xiaoying He
Nanjing Tech University
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Yan Li
Nanjing Tech University
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Honghua Jia
Nanjing Tech University
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Ming Yan
Nanjing Tech University
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Abstract

Abstract: 2-O-α-D-Glucopyranosyl-L-ascorbic acid (AA-2G) is an important industrial derivative of L-ascorbic acid (AA), which has the distinct advantages of non-reducibility, antioxidation, and reproducible decomposition into L-ascorbic acid and glucose. Enzymatic synthesis is a preferred method for AA-2G production over alternative chemical synthesis owing to the regioselective glycosylation reaction. α-Glucosidase, an enzyme classed into O- glycoside hydrolases, may be used in glycosylation reactions to synthesize AA-2G. Here, one α-glucosidase from Oryza sativa (rAGL) was recombinantly produced in Pichia pastoris GS115 and used for biosynthesis of AA-2G with few intermediates and byproducts. The extracellular rAGL reached 9.11 U/mL after fed-batch cultivation for 102 h in a 5-L fermenter. The specific activity of purified rAGL is 49.83 U/mg at 37 °C and pH 4.0. The optimal temperature of rAGL was 65 °C, and it was stable below 55 °C. rAGL was active over the range of pH 3.0–7.0, with the maximal activity at pH 4.0. Under the condition of 37 °C , pH 4.0, equimolar maltose and AA·Na, 8.7±0.4 g/L of AA-2G was synthesized by rAGL. These studies lay the basis for the industrial application of recombinant α-glucosidase. Keywords: α-Glucosidase; Oryza sativa; 2-O-α-D-glucopyranosyl-L-ascorbic acid; Transglycosylation; Pichia pastoris

Peer review status:IN REVISION

16 Apr 2021Submitted to Biotechnology Journal
16 Apr 2021Submission Checks Completed
16 Apr 2021Assigned to Editor
14 May 2021Reviewer(s) Assigned
27 May 2021Editorial Decision: Revise Minor
10 Jun 20211st Revision Received
10 Jun 2021Reviewer(s) Assigned
10 Jun 2021Submission Checks Completed
10 Jun 2021Assigned to Editor