Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Chemical Engineering, Nanjing Forestry University, Nanjing, 210037, People's Republic of China.
School of Grain Science and Technology, Jiangsu University of Science and Technology, Zhenjiang, 212003, People's Republic of China.
Microb Cell Fact. 2022 Sep 8;21(1):184. doi: 10.1186/s12934-022-01907-0.
Efficient upgrading of inferior agro-industrial resources and production of bio-based chemicals through a simple and environmentally friendly biotechnological approach is interesting Lactobionic acid is a versatile aldonic acid obtained from the oxidation of lactose. Several microorganisms have been used to produce lactobionic acid from lactose and whey. However, the lactobionic acid production titer and productivity should be further improved to compete with other methods.
In this study, a new strain, Pseudomonas fragi NL20W, was screened as an outstanding biocatalyst for efficient utilization of waste whey to produce lactobionic acid. After systematic optimization of biocatalytic reactions, the lactobionic acid productivity from lactose increased from 3.01 g/L/h to 6.38 g/L/h in the flask. In batch fermentation using a 3 L bioreactor, the lactobionic acid productivity from whey powder containing 300 g/L lactose reached 3.09 g/L/h with the yield of 100%. Based on whole genome sequencing, a novel glucose dehydrogenase (GDH1) was determined as a lactose-oxidizing enzyme. Heterologous expression the enzyme GDH1 into P. putida KT2440 increased the lactobionic acid yield by 486.1%.
This study made significant progress both in improving lactobionic acid titer and productivity, and the lactobionic acid productivity from waste whey is superior to the ever reports. This study also revealed a new kind of aldose-oxidizing enzyme for lactose oxidation using P. fragi NL20W for the first time, which laid the foundation for further enhance lactobionic acid production by metabolic engineering.
通过简单环保的生物技术方法,高效升级劣质农业工业资源并生产生物基化学品是很有趣的。乳寡糖是一种从乳糖氧化得到的多功能醛糖。已经有几种微生物被用于从乳糖和乳清生产乳寡糖。然而,为了与其他方法竞争,乳寡糖的生产滴度和生产率还需要进一步提高。
在这项研究中,筛选出一种新的菌株——弗氏假单胞菌 NL20W,作为一种高效利用废乳清生产乳寡糖的优秀生物催化剂。在生物催化反应的系统优化后,乳糖的乳寡糖生产率从摇瓶中的 3.01 g/L/h 提高到 6.38 g/L/h。在使用 3 L 生物反应器的分批发酵中,从含有 300 g/L 乳糖的乳清粉中生产乳寡糖的生产率达到 3.09 g/L/h,产率为 100%。基于全基因组测序,确定了一种新型葡萄糖脱氢酶(GDH1)为乳糖氧化酶。将酶 GDH1 异源表达到 P. putida KT2440 中,使乳寡糖的产率提高了 486.1%。
本研究在提高乳寡糖的滴度和生产率方面取得了重大进展,且从废乳清中生产乳寡糖的生产率优于以往的报道。本研究还首次揭示了一种利用弗氏假单胞菌 NL20W 氧化乳糖的新型醛糖氧化酶,为通过代谢工程进一步提高乳寡糖产量奠定了基础。