Effect of exogenous gene expression on sugar alcohol metabolism of Yarrowia lipolytica
ZHOU Yunqi1, LI Hongjiao1, LI Kewen2, GUO Bei3, PEI Jiangsen1*
1(China National Research Institute of Food and Fermentation Industries, Beijing 100015, China) 2(Bao Ling Bao Biology Company Limited, Yucheng 251200, China) 3(College of Bioscience and Resources Environment, Beijing University of Agriculture, Beijing 102206, China)
Abstract: Yarrowia lipolytica is a widely used eukaryotic expression host and a major strain for erythritol fermentation in China. In this research, a putative gene with sugar phosphorylation function cloned from Zygoascus hellenicus was constructed into an expression vector and transformed into Y. lipolytica. The effect of the recombinant strain on substrate utilization and conversion related to erythritol synthesis was investigated. The results showed that in the shake flask fermentation, different assimilation of erythritol and ribose was observed in the recombinant Y. lipolytica compared with the original strain. With glucose and erythritol/ribose in the medium, the assimilation ability of the recombinant strain was at least twice that of the original strain. This functional gene was a new gene with the ability to regulate erythritol synthesis. The treatment experiment of erythritol waste liquor showed that the recombinant strain could assimilate various sugar alcohols for cell growth compared with the control strain, and change the composition of the crystallization waste liquor to reduce the viscosity and maltose concentration, providing a new solution for the comprehensive utilization of industrial waste liquor.
[1] 肖素荣, 李京东.赤藓糖醇的特性及应用[J].中国食物与营养, 2008(5):26-28. XIAO S R, LI J D.Characteristics and application of erythritol[J].Food and Nutrition in China, 2008(5):26-28. [2] WEN H, TANG B, STEWART A J, et al.Erythritol attenuates postprandial blood glucose by inhibiting α-glucosidase[J].Journal of Agricultural & Food Chemistry, 2018,66(6):1 401-1 407. [3] REGNAT K, MACH R L, MACH-AIGNER A R.Erythritol as sweetener-wherefrom and whereto?[J].Applied Microbiology & Biotechnology, 2018,102(2):587-595. [4] HOOTMAN K C, TREZZI J, KRAEMER L, et al.Erythritol is a pentose-phosphate pathway metabolite and associated with adiposity gain in young adults[J].Proceedings of the National Academy of Sciences of the United States of America, 2017,114(21):E4 233-E4 240. [5] 高蕾蕾, 刘峰, 栾庆民, 等.赤藓糖醇生产与应用研究进展[J].精细与专用化学品, 2020,28(3):1-4. GAO L L, LIU F, LUAN Q M, et al.The research progress in erythritol production technology and application[J].Fine and Specialty Chemicals, 2020,28(3):1-4. [6] 徐莹, 李景军, 何国庆.赤藓糖醇研究进展及在食品中的应用[J].中国食品添加剂, 2005(3):92-95;54. XU Y, LI J J, HE G Q.Research progress on erythritol and application in food science[J].China Food Additives, 2005(3):92-95;54. [7] KOBAYASHI Y, IWATA H, MIZUSHIMA D, et al.Erythritol production by Moniliella megachiliensis using nonrefined glycerol waste as carbon source[J].Letters in Applied Microbiology, 2015,60(5):475-480. [8] WANG S Q, WANG H W, LYU J Y, et al.Highly efficient erythritol recovery from waste erythritol mother liquor by a yeast-mediated biorefinery process[J].Journal of Agricultural and Food Chemistry, 2017,65(50):11 020-11 028. [9] 裴疆森, 黄玲, 张露, 等.解脂亚罗酵母赤藓糖醇发酵过程的研究[J].食品与发酵工业, 2015,41(4):28-33. PEI J S, HUANG L, ZHANG L, et al.Erythritol production with Yarrowia lipolitica[J].Food and Fermentation Industries, 2015,41(4):28-33. [10] RZECHONEK D A, DOBROWOLSKI A, RYMOWICZ W, et al.Recent advances in biological production of erythritol[J].Critical Reviews in Biotechnology, 2018,38(4):620-633. [11] 刘俊梅, 王辉, 李琢伟, 等.赤藓糖醇合成途径及关键酶的研究进展[J].粮油加工(电子版), 2015(5):58-61;65. LIU J M, WANG H, LI Z W, et al.Research progress of erythritol biosynthetic pathway and key enzymes[J].Cereals and Oils Processing(Electronic Version), 2015(5):58-61;65. [12] 程磊, 宗朕, 陈卓静, 等.酵母菌发酵生产赤藓糖醇的研究进展[J].中国调味品, 2018,43(12):181-186. CHENG L, ZONG Z, CHEN Z J, et al.Research progress on erythritol production by yeast fermentation[J].China Condiment, 2018,43(12):181-186. [13] 殷晓霞. 代谢工程改造解脂亚洛酵母产α-酮戊二酸[D].无锡:江南大学, 2012. YIN X X.Metabolic engineering of Yarrowia lipolytica for α-ketoglutarate production[D].Wuxi:Jiangnan University, 2012. [14] MADZAK C, TRÉTON B, BLANCHIN-ROLAND S.Strong hybrid promoters and integrative expression/secretion vectors for quasi-constitutive expression of heterologous proteins in the yeast Yarrowia lipolytica[J].Journal of Molecular Microbiology and Biotechnology, 2000,2(2):207. [15] MADZAK C.Yarrowia lipolytica:Recent achievements in heterologous protein expression and pathway engineering[J].Applied Microbiology and Biotechnology, 2015,99(11):4 559-4 577. [16] 赵鹤云, 黄瑛, 杨江科, 等.解脂耶氏酵母表达系统研究进展[J].生物加工过程, 2008(3):10-16. ZHAO H Y, HUANG Y, YANG J K, et al.Review of Yarrowia lipolytica expression system[J].Chinese Journal of Bioprocess Engineering, 2008(3):10-16. [17] MARKHAM K A, ALPER H S.Synthetic biology expands the industrial potential of Yarrowia lipolytica[J].Trends in Biotechnology, 2018,36(10):1 085-1 095. [18] MADZAK C, GAILLARDIN C, BECKERICH J.Heterologous protein expression and secretion in the non-conventional yeast Yarrowia lipolytica:A review[J].Journal of Biotechnology, 2004,109(1-2):63-81. [19] JANEK T, DOBROWOLSKI A, BIEGALSKA A, et al.Characterization of erythrose reductase from Yarrowia lipolytica and its influence on erythritol synthesis[J].Microbial Cell Factories, 2017,16(118).DOI:10.1186/s12934-017-0733-6. [20] CARLYA F, VANDERMIES M, TELEK S, et al.Enhancing erythritol productivity in Yarrowia lipolytica using metabolic engineering[J].Metabolic Engineering, 2017,42:19-24. [21] CARLY F, STEELS S, TELEK S, et al.Identification and characterization of EYD1, encoding an erythritol dehydrogenase in Yarrowia lipolytica and its application to bioconvert erythritol into erythrulose[J].Bioresource Technology, 2018,247:963-969. [22] 田强, 秦海青, 邱学良, 等.Yarrowia lipolytica酵母产赤藓糖醇的工艺研究[J].中国食品添加剂, 2015(9):121-124. TIAN Q, QIN H Q, QIU X L, et al.Research on the process for erythritol production by Yarrowia lipolytica yeast[J].China Food Additives, 2015(9):121-124. [23] 田生礼, 梁秀怡, 梁志成, 等.T载体的应用及研究进展[J].科学技术与工程, 2014, 14(30):102-108. TIAN S L, LIANG X Y, LIANG Z C, et al.Application and research progress of T vector[J].Science Technology and Engineering, 2014,14(30):102-108. [24] 楼佳佳, 韦柳静, 花强.解脂耶氏酵母异源合成番茄红素的初步研究[J].华东理工大学学报(自然科学版), 2015,41(3):314-320. LOU J J, WEI L J, HUA Q.Metabolic engineering study of heterologous production of lycopene in Yarrowia lipolytica[J].Journal of East China University of Science and Technology (Natural Science Edition), 2015,41(3):314-320. [25] 赵祥颖, 张立鹤, 杨丽萍, 等.赤藓糖醇生产母液中糖醇组分的定性鉴定[J].食品与发酵工业, 2014,40(10):200-204. ZHAO X Y, ZHANG L H, YANG L P, et al.Identification of the components in the erythritol mother liquor[J].Food and Fermentation Industries, 2014,40(10):200-204. [26] NAKAGAWA Y, KASUMI T, OGIHARA J, et al.Erythritol:Another C4 platform chemical in biomass refinery[J].ACS Omega, 2020,5(6):2 520-2 530.
2021, Vol. 47 
