{"title":"左旋葡聚糖激酶活性和转运能力限制了左旋葡聚糖在酿酒酵母中的利用。","authors":"Mengdan Yang, Tiandi Wei, Kai Wang, Liqun Jiang, Dihao Zeng, Xinhua Sun, Weifeng Liu, Yu Shen","doi":"10.1186/s13068-022-02195-x","DOIUrl":null,"url":null,"abstract":"<p><p>Manufacturing fuels and chemicals from cellulose materials is a promising strategy to achieve carbon neutralization goals. In addition to the commonly used enzymatic hydrolysis by cellulase, rapid pyrolysis is another way to degrade cellulose. The sugar obtained by fast pyrolysis is not glucose, but rather its isomer, levoglucosan (LG). Here, we revealed that both levoglucosan kinase activity and the transportation of levoglucosan are bottlenecks for LG utilization in Saccharomyces cerevisiae, a widely used cell factory. We revealed that among six heterologous proteins that had levoglucosan kinase activity, the 1,6-anhydro-N-acetylmuramic acid kinase from Rhodotorula toruloides was the best choice to construct levoglucosan-utilizing S. cerevisiae strain. Furthermore, we revealed that the amino acid residue Q341 and W455, which were located in the middle of the transport channel closer to the exit, are the sterically hindered barrier to levoglucosan transportation in Gal2p, a hexose transporter. The engineered yeast strain expressing the genes encoding the 1,6-anhydro-N-acetylmuramic acid kinase from R. toruloides and transporter mutant Gal2p<sup>Q341A</sup> or Gal2p<sup>W455A</sup> consumed ~ 4.2 g L<sup>-1</sup> LG in 48 h, which is the fastest LG-utilizing S. cerevisiae strain to date.</p>","PeriodicalId":9125,"journal":{"name":"Biotechnology for Biofuels and Bioproducts","volume":" ","pages":"94"},"PeriodicalIF":0.0000,"publicationDate":"2022-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9476349/pdf/","citationCount":"0","resultStr":"{\"title\":\"Both levoglucosan kinase activity and transport capacity limit the utilization of levoglucosan in Saccharomyces cerevisiae.\",\"authors\":\"Mengdan Yang, Tiandi Wei, Kai Wang, Liqun Jiang, Dihao Zeng, Xinhua Sun, Weifeng Liu, Yu Shen\",\"doi\":\"10.1186/s13068-022-02195-x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Manufacturing fuels and chemicals from cellulose materials is a promising strategy to achieve carbon neutralization goals. In addition to the commonly used enzymatic hydrolysis by cellulase, rapid pyrolysis is another way to degrade cellulose. The sugar obtained by fast pyrolysis is not glucose, but rather its isomer, levoglucosan (LG). Here, we revealed that both levoglucosan kinase activity and the transportation of levoglucosan are bottlenecks for LG utilization in Saccharomyces cerevisiae, a widely used cell factory. We revealed that among six heterologous proteins that had levoglucosan kinase activity, the 1,6-anhydro-N-acetylmuramic acid kinase from Rhodotorula toruloides was the best choice to construct levoglucosan-utilizing S. cerevisiae strain. Furthermore, we revealed that the amino acid residue Q341 and W455, which were located in the middle of the transport channel closer to the exit, are the sterically hindered barrier to levoglucosan transportation in Gal2p, a hexose transporter. The engineered yeast strain expressing the genes encoding the 1,6-anhydro-N-acetylmuramic acid kinase from R. toruloides and transporter mutant Gal2p<sup>Q341A</sup> or Gal2p<sup>W455A</sup> consumed ~ 4.2 g L<sup>-1</sup> LG in 48 h, which is the fastest LG-utilizing S. cerevisiae strain to date.</p>\",\"PeriodicalId\":9125,\"journal\":{\"name\":\"Biotechnology for Biofuels and Bioproducts\",\"volume\":\" \",\"pages\":\"94\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2022-09-14\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9476349/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biotechnology for Biofuels and Bioproducts\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1186/s13068-022-02195-x\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology for Biofuels and Bioproducts","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1186/s13068-022-02195-x","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
从纤维素材料制造燃料和化学品是实现碳中和目标的一个有前途的策略。除了常用的纤维素酶水解法外,快速热解法是降解纤维素的另一种方法。快速热解得到的糖不是葡萄糖,而是它的异构体,左旋葡聚糖(LG)。在这里,我们发现左旋葡聚糖激酶活性和左旋葡聚糖的运输是酿酒酵母利用LG的瓶颈,酿酒酵母是广泛使用的细胞工厂。结果表明,在具有左旋葡聚糖激酶活性的6个异源蛋白中,以红酵母1,6-无氢- n-乙酰氨基酸激酶为载体构建酿酒酵母左旋葡聚糖菌株的最佳选择。此外,我们发现位于运输通道中间靠近出口的氨基酸残基Q341和W455是Gal2p(一种己糖转运体)中左旋葡聚糖运输的空间障碍屏障。从toruloides和转运体突变体Gal2pQ341A或Gal2pW455A中表达1,6-无氢- n -乙酰氨基酸激酶编码基因的工程酵母菌在48 h内消耗了4.2 g L-1 LG,是迄今为止最快的利用LG的酿酒酵母菌株。
Both levoglucosan kinase activity and transport capacity limit the utilization of levoglucosan in Saccharomyces cerevisiae.
Manufacturing fuels and chemicals from cellulose materials is a promising strategy to achieve carbon neutralization goals. In addition to the commonly used enzymatic hydrolysis by cellulase, rapid pyrolysis is another way to degrade cellulose. The sugar obtained by fast pyrolysis is not glucose, but rather its isomer, levoglucosan (LG). Here, we revealed that both levoglucosan kinase activity and the transportation of levoglucosan are bottlenecks for LG utilization in Saccharomyces cerevisiae, a widely used cell factory. We revealed that among six heterologous proteins that had levoglucosan kinase activity, the 1,6-anhydro-N-acetylmuramic acid kinase from Rhodotorula toruloides was the best choice to construct levoglucosan-utilizing S. cerevisiae strain. Furthermore, we revealed that the amino acid residue Q341 and W455, which were located in the middle of the transport channel closer to the exit, are the sterically hindered barrier to levoglucosan transportation in Gal2p, a hexose transporter. The engineered yeast strain expressing the genes encoding the 1,6-anhydro-N-acetylmuramic acid kinase from R. toruloides and transporter mutant Gal2pQ341A or Gal2pW455A consumed ~ 4.2 g L-1 LG in 48 h, which is the fastest LG-utilizing S. cerevisiae strain to date.