Whole-cell catalytic synthesis of 2-O-α-glucopyranosyl-l-ascorbic acid by sucrose phosphorylase from Bifidobacterium breve via a batch-feeding strategy. (January 2022)

• Record Type: Journal Article
• Authors: Zhou, Yaoyao; Gan, Tian; Jiang, Ruini; Chen, Hanchi; Ma, Zhi; Lu, Yuele; Zhu, Linjiang; Chen, Xiaolong
• Is Part Of: Process biochemistry. Volume 112(2022)
• Page Start: 27
• Page End: 34
• Publication Date: 2022-01
• Subjects: SPase sucrose phosphorylase -- Bbr Bifidobacterium breve -- Blo Bifidobacterium longum -- Lac Lactobacillus acidophilus -- Lre Lactobacillus reuteri -- Ljo Lactobacillus johnsonii -- Bla Bifidobacterium lactis -- Lem Leuconostoc mesenteroides -- T/H transglycosylation/hydrolysis; Sucrose phosphorylase -- l-ascorbic acid 2-O-α-d-glucoside -- Whole-cell catalysis -- Batch-feeding; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2021.11.023
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Three typical activities of eight sucrose phosphorylases (SPase) were compared. Fructose remarkably inhibits the glycosylation of l -ascorbic acid by SPase. Whole-cell transformation is effective for the glycosylation of l -ascorbic acid. Sucrose feeding strategy increased molar conversion rate of l -ascorbic acid. Abstract: l -ascorbic acid 2- O - α -d -glucoside (AA-2G), a highly stabilized vitamin C derivative, has been widely applied in cosmetics, foods, and pharmaceuticals. Sucrose phosphorylase (SPase) was recently reported to be effective for producing AA-2G. In this study, SPase from Bifidobacterium breve (BbrSPase) was identified as a new AA-glycosylating tool with high activity and good thermostability. The optimal conditions for AA glycosylation are pH 5.5 and 55 °C. The K m and k c a t when sucrose was applied as a glycosyl donor were 2.72 ± 0.49 mM and 9.70 ± 0.10 s −1, respectively. The transglycosylation activity was seriously inhibited by fructose, which caused a sharp decline in the glycosylation rate but did not necessarily stop the yield of AA-2G due to the high affinity to sucrose. Therefore, to attenuate the inhibition of fructose, a new sucrose batch-feeding strategy was developed for the effective production of AA-2G. The maximum titer was 185 g/L within 72 h. The molar conversion rate of l -AA reached 50.2 %, which is the highest conversion rate to our knowledge.