食品与发酵工业2024，Vol.50Issue(16)：1-9,9.DOI:10.13995/j.cnki.11-1802/ts.037798
重组地衣芽孢杆菌全细胞转化法制备D-阿洛酮糖
Whole-cell biosynthesis of D-allulose by recombinant Bacillus licheniformis
摘要
Abstract
D-allulose(D-psicose)is a low-calorie functional sugar,which has many functions such as sucrose substitute,participat-ing in Maillard reaction,improving food gel process,etc.One of the methods of preparing D-allulose,biosynthesis,has many advantages such as simple purification steps,high product concentration,good environmental compatibility,etc.Therefore,the preparation of D-allu-lose by biosynthesis has become a research hotspot.In this study,Bacillus licheniformis is used to heterologously express D-psicose 3-epi-merase(DPEase)from Ruminiclostridium cellulolyticum H10 for the first time,which opens up a new expression vector pathway for D-al-lulose biosynthesis.Firstly,different promoters were selected to mediate the expression of DPEase,and the recombinant strain BL1 with the best expression effect was selected to optimize the whole-cell transformation and fermentation conditions.The optimized recombinant bacterial whole-cell transformation conditions were that the temperature was 65 ℃,the cell OD600 of the reaction system was 2,the reac-tion time was 10 minutes,and the substrate D-fructose was 100 g/L,which was utilized to explore the fermentation medium conditions,which was that the carbon source was 75 g/L sucrose,the initial pH was 7.5,and the temperature was 37 ℃.Finally,the whole-cell bio-synthesis transformation method was used,which was that the temperature was 65 ℃,D-fructose was 500 g/L,the cell OD600 was 30,and the reaction time was 20 minutes,the conversion rate was achieved at 30.3％,the D-allulose was converted to 120 g/L,and unit enzyme activity reached 33.3 U/mL.The conversion rate was increased to 69.8％by adding boric acid with a molar mass ratio of 0.4 to fructose in four stages of the reaction system.This study provides a certain reference value for the industrial production of D-allulose.关键词
D-阿洛酮糖/地衣芽孢杆菌/全细胞转化/D-阿洛酮糖3-差向异构酶/异源表达/启动子Key words
D-allulose/Bacillus licheniformis/whole-cell catalysis/D-psicose 3-epimerase/heterologous expression/promoter引用本文复制引用
魏雨,李由然,石贵阳..重组地衣芽孢杆菌全细胞转化法制备D-阿洛酮糖[J].食品与发酵工业,2024,50(16):1-9,9.基金项目
国家重点研发计划项目(2020YFA0907704) （2020YFA0907704）
国家自然科学基金项目(32172174) （32172174）
