Study on Transglucosylation Properties of Amylosucrase from Xanthomonas campestris pv. Campestris and Its Application in the Production of α-Arbutin

Abstract

α-Arbutin (4-hydroquinone-α-D-glucopyranoside), an effective skin-lightening agent due to its considerable inhibitory effect on human tyrosinase activity, is widely used in the pharmaceutical and cosmetic industries. Recently, α-arbutin was prepared through transglucosylation of hydroquinone using microbial glycosyltransferases as catalysts. However, the low yield and prolonged reaction time of the biotransformation process of α-arbutin production limited its industrial application. In this work, an amylosucrase (ASase) from Xanthomonas campestris pv. campestris str. ATCC 33913 (XcAS) was expressed efficiently in Escherichia coli JM109. The catalytic property of the purified XcAS for the synthesis of α-arbutin was tested. The recombinant strain was applied for highly efficient synthesis of α-arbutin using sucrose and hydroquinone as glucosyl donor and acceptor, respectively. By optimizing the biotransformation conditions and applying a fed-batch strategy, the final production yield and conversion rate of α-arbutin reached 60.9 g/L and 95.5%, respectively, which is the highest reported yield by engineered strains. Compared to the highest reported value (<1.4 g/L/h), our productivity (7.6 g/L/h) was improved more than five-fold. This work represents an efficient and rapid method for α-arbutin production with potential industrial applications.

Optimization of whole-cell biotransformation for scale-up production of α-arbutin from hydroquinone by the use of recombinant Escherichia coli

“…Whereas, the molar conversion rate was almost equivalent to that in laboratory scale. The yield was still in the highest level by comparing to previous report (Yang et al 2019; Zhu et al 2018). Therefore, the scale-up production of α-arbutin by batch-feeding whole-cell biotransformation is satisfactory.…”

“…Its activity declined by 30–40% due to the lack of kanamycin in comparison with the cells cultured in 2.5 L fermenter. The final concentrations of 375–397 mM (102–108 g/L), molar conversion rate (~ 95%) and productivity (4.53–4.91 kg kL −1 h −1 ) were satisfactory, which were much higher than those of scale-up production by the cells of Xanthomonas BT-112 (Wei et al 2016) and recombinant E. coli cells (Yang et al 2019). Therefore, batch-feeding whole-cell biotransformation by recombinant E. coli cells expressing Amy-1 is effective for industrial production of α-arbutin.…”

“…Subsequently, sucrose phosphorylase from Leuconostoc mesenteroides and α-glucosidase from Xanthomonas campestris were reported with a higher molar conversion rate of 60% and 93%, respectively, but only a low concentration of α-arbutin (~ 10 g/L) was obtained (Kitao and Sekine 1994; Kurosu et al 2002; Sato et al 2012). In recent years, various enzymes were newly identified with HQ glycosylation activity including dextransucrase from L. mesenteroides , cyclodextrin glucanotransferase from Thermoanaerobacter sp., amylosucrases from Cellulomonas carboniz , Deinococcus geothermalis , and X. campestris (Mathew and Adlercreutz 2013; Seo et al 2009; Yang et al 2019; Yu et al 2018). Amylosucrase displayed a highest HQ glycosylation activity among these enzymes, for example, obtaining > 90% of molar conversion rate and 60 g/L of concentration (Yang et al 2019).…”

“…In recent years, various enzymes were newly identified with HQ glycosylation activity including dextransucrase from L. mesenteroides , cyclodextrin glucanotransferase from Thermoanaerobacter sp., amylosucrases from Cellulomonas carboniz , Deinococcus geothermalis , and X. campestris (Mathew and Adlercreutz 2013; Seo et al 2009; Yang et al 2019; Yu et al 2018). Amylosucrase displayed a highest HQ glycosylation activity among these enzymes, for example, obtaining > 90% of molar conversion rate and 60 g/L of concentration (Yang et al 2019). We also recently identified the amylosucrase (Amy-1) from Xanthomonas campestris with > 95% molar conversion rate and > 80 g/L of α-arbutin concentration (Fig.…”

Functionalized chitosan nanoparticles for cutaneous delivery of a skin whitening agent: an approach to clinically augment the therapeutic efficacy for melasma treatment

“…It is reported that arbutin's activity comes from its structural homologies with its substrate (tyrosine), which causes the competitive suppression of tyrosinase catalytic activity (Sugimoto et al., 2004 ). A number of studies have revealed that alpha-arbutin (α-arbutin) (or 4-hydroxyphenyl alpha-glucopyranoside) shows more substantial inhibitory potential on human tyrosinase, reaching ten times greater effect compared to beta-arbutin, which promotes its efficacy in attaining the required whitening effect on the skin (Yang et al., 2018 ).…”