Development of an enzymatic synthesis approach to produce phloridzin using Malus x domestica glycosyltransferase in engineered Pichia pastoris GS115. (August 2017)

• Record Type: Journal Article
• Authors: Zhang, Ting-Jing; Liang, Jian-Qiang; Wei, Xin-Yuan; Wang, Pan-Xue; Xu, Ying; Pen, Shuai; Fan, Ming-Tao
• Is Part Of: Process biochemistry. Volume 59:Part B(2017)
• Page Start: 187
• Page End: 193
• Publication Date: 2017-08
• Subjects: Phloridzin -- Phloretin-2′-O-glycosyltransferase -- Phloretin -- Engineered P. pastoris GS115 -- Purification; 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.2017.05.009
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Enzymatic biocatalyst approach was developed for the production of phloridzin. The conditions for the production of phloridzin were optimized by RSM. Analytical grade of phloridzin was efficiently obtained. Phloridzin purity and yield obtained with methods developed are the highest reported to date. Abstract: Phloridzin has been an attractive target for the development of preparation methodologies due to its various applications as pharmaceutical ingredient and food additive. In this study, an enzymatic synthesis approach using engineered P. pastoris GS115 was developed to produce phloridzin. Four factors, identifying as key factors affecting the phloridzin yield, were optimized by response surface methodology as follows: induction time 121.8 h, pH 7.0, methanol 0.75%, and a rotation speed 194 rpm. The maximal yield of phloridzin reached 41.59 mg/L (95.31 μM) and the specific bioconversion rate of substrate was up to 98.50% with 1L working volume in a 5L bioreactor under the optimal conditions. Phloridzin with purity of 93.98% and recovery of 78.14% was achieved just using affinity chromatography and preparative high performance liquid chromatography ( Pre -HPLC). These results confirm the enzymatic synthesis developed provides an efficient alternative to traditional approach for the preparation of phloridzin in vitro .