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For each citation that was shared on social media (LinkedIn, Facebook, or Twitter) with the “@GenScript” tag, the author will be rewarded with a $10 Amazon gift card or 2,000 GS points.Preparation of low-molecular-weight citrus pectin by recombinant Bacillus subtilis pectate lyase and promotion of growth of Bifidobacterium longum
Catalysis Communications. 2018;Products/Services Used Details Operation Recombinant Proteins Bacillus subtilis JL-13 strain, which produces pectate lyase, was isolated from the soil of Hangzhou Botanical Garden in our previous study [10]. The fusion cold shock expression vector, pCold TF, was purchased from Takara Biotechnology Co., Ltd. (Dalian, China). Medium components were obtained from Oxoid. Citrus pectin, D-(+)-galacturonic acid, polygalacturonate, and bovine serum albumin (BSA) were acquired from Sigma-Aldrich (Shanghai) Trading Co., Ltd. (Shanghai, China). High-affinity Ni-charged resin was provided by GenScript Biotechnology Co., Ltd. (Nanjing, China). All other chemicals used in this study were of analytical grade. Get A Quote Abstract
pel gene (GenBank: KM244046.1), which encodes Bacillus subtilis pectate lyase (PelB), was high-level expressed in Escherichia coli BL21 (DE3) and the recombinant enzyme was named rePelB. After induction by IPTG at 15 °C for 24 h, specific activity of rePelB in culture supernatant of pCpel-6 reached 197.4 ± 5.20 U/mg. The optima of rePelB were 60 °C and pH 5.0, respectively. Hydrolysis of citrus pectin by rePelB in 50 L grass reactor was optimized by implementing RSM. HPLC analysis revealed molecular mass of citrus pectin and obtained LCP was 290 and 5.8 kDa, respectively. LCP can significantly promote growth of Bifidobacterium longum.Keywords
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