Improvement of long chain dicarboxylic acid production by fortifying omega-oxidation, excretion of products and lipase production in yeast

通过强化酵母中的欧米伽氧化、产物排泄和脂肪酶生产来改善长链二羧酸的生产

• 批准号: 12556011
• 负责人: OHTA Akinori
• 金额: $ 8.64万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12556011/
• 关键词: long chain dicarboxylic acid; Yeast; Candida maltosa; cytochrome P450; oxidation of n-alkanes; ω oxidation; ALK genes; ABC transporter; n-アルカン; 脂肪酸のω酸化; n-アルカンの

Result of this research is constituted of following two parts.From comparison of wild type and the long chain dicarboxylic acid (DCA)-producing mutant of Candida maltosa, we found that ALK5 gene that is active in ω-oxidation of fatty acids was highly produced in the mutant, which is presumed to be important for DCA production. Improvement of ALK5 expression by alteration of promoter structure should be useful try in future.We also found that a gne encoding one of ABC transporters was strongly induced at the time of DCA production in the mutant.. The identified gene was a homolog of CDR1 of Candida albicance, which confers cycloheximide resistance to this pathogenic yeast. Further investigation is necessary for full elucidation of its function in DCA production in C. maltosa.

本研究的结果主要包括以下两个部分：通过对野生型和长链二羧酸（DCA）产生菌的比较，发现具有ω-氧化脂肪酸活性的ALK 5基因在突变株中大量表达，推测其对DCA的产生具有重要作用。通过改变启动子结构来提高ALK 5的表达应该是未来有用的尝试。我们还发现，在突变体中产生DCA时，编码ABC转运蛋白之一的基因被强烈诱导。鉴定的基因是白色念珠菌的CDR 1的同源物，其赋予放线菌酮抗性到该致病性酵母。进一步的研究是必要的，以充分阐明其功能，在DCA生产的C。麦芽糖。

项目成果

T.Sumita, T.Iida, et al.: "Y1ALK1 encoding the cytochrome P45OALK1 in Yarrowia lipolytica is transcriptionally induced by n-decane through two distinct cis-elements on its promoter"Biochem. Biophys. Res. Commun.. 294. 1071-1078 (2002)

T.Sumita、T.Iida 等人：“解脂耶氏酵母中编码细胞色素 P45OALK1 的 Y1ALK1 由正癸烷通过其启动子上的两个不同顺式元件进行转录诱导”Biochem.

T. Zimmer, A. Ogura et al.: "Gene regulation in response to over expression of cytochrome P450 and proliferation of the endoplasmic reticulum in Saccharomyces cerevisiae"Biosci. Biotechnol. Biochem.. 64. 1930-1936 (2000)

T. Zimmer、A. Ogura 等人：“酿酒酵母中细胞色素 P450 过度表达和内质网增殖的基因调控”Biosci。

T.Sumita, T.Iida, et al.: "Cloning and characterization of Yarrowia lipolytica PEX10 : Its deficiency represses the expression of n-alkane-inducible Y1ALK1 encoding cytochrome P45OALK1"FEMS Microbiolgy Letters. 214. 31-38 (2002)

T.Sumita、T.Iida 等人：“解脂耶氏酵母 PEX10 的克隆和表征：其缺陷会抑制编码细胞色素 P45OALK1 的正烷烃诱导型 Y1ALK1 的表达”FEMS 微生物学快报。

T. lida, T. Sumita, et al.: "The cytochrome P450ALK multigene family of an n-alkane-assimilating yeast, Yarrowia lipolytica: characterization of genes coding for new CYP52 family members"Yeast. 16. 1077-1067 (2000)

T. lita、T. Sumita 等人：“正烷烃同化酵母 Yarrowia lipolytica 的细胞色素 P450ALK 多基因家族：编码新 CYP52 家族成员的基因的特征”酵母。

T.Zimrner, A.Ogura et al.: "Gene regulation in response to over expression of cytochrome P450 and proliferation of the endoplasmic reticulum in Saccharomyces cerervisiae"Biosci. Biotechnol. Biochem.. 64. 1930-1936 (2000)

T.Zimrner、A.Ogura 等人：“酿酒酵母中细胞色素 P450 过度表达和内质网增殖的基因调控”Biosci。