Engineering Corynebacterium glutamicum Mutants for 3-Methyl-1-butanol Production
- 51 Downloads
3-Methyl-1-butanol (3MB) is a promising biofuel that can be produced from 2-ketoisocaproate via the common l-leucine biosynthesis pathway. Corynebacterium glutamicum was chosen as a host bacterium because of its strong resistance to isobutanol. In the current study, several strategies were designed to overproduce 3MB in C. glutamicum through a non-fermentation pathway. The engineered C. glutamicum mutant was obtained by silencing the pyruvate dehydrogenase gene complex (aceE) and deleting the lactic dehydrogenase gene (ldh), followed by mutagenesis with diethyl sulfate (DES) and selection with Fmoc-3-4-thiazolyl-l-alanine (FTA). The mutant could produce 659 mg/L of 3MB after 12 h of incubation. To facilitate carbon flux to 3MB biosynthesis, the engineered recombinant was also constructed without branched-chain acid aminotransferase (ilvE) activity by deleting the ilvE gene. This recombinant could produce 697 mg/L of 3MB after 12 h of incubation.
Keywords3-Methyl-1-butanol l-Leucine Corynebacterium glutanicum Mutation breeding Non-fermentation pathway
This research was financially supported by National Natural Science Foundation of China (21506215 and 21811530626), Municipal Science and Technology Program of Guangzhou (China) (201804010081).
- Li S (2011) Biofuel as an alternative to oil—current industry progress and sustainable development. Eng Sci 13:50–56Google Scholar
- Liu Y, Liu H, Zhang J, Cheng K, Chen Z (2008) Research progress in new biofuel butanol Xiandai Huagong/modern. Chem Ind 28(28–31):33Google Scholar
- Patek M, Krumbach K, Eggeling L, Sahm H (1994) Leucine synthesis in corynebacterium-glutamicum— enzyme-activities, structure of leua, and effect of leua inactivation on lysine synthesis. Appl Environ Microbiol 60:133–140Google Scholar
- Schafer A, Tauch A, Jager W, Kalinowski J, Thierbach G, Puhler A (1994) Small mobilizable multipurpose cloning vectors derived from the Escherichia-coli plasmids PK18 and PK19—selection of defined deletions in the chromosome of corynebacterium-glutamicum. Gene 145:69–73. https://doi.org/10.1016/0378-1119(94)90324-7 CrossRefGoogle Scholar
- Sikkema J, Debont JAM, Poolman B (1995) Mechanisms of membrane toxicity of hydrocarbons. Microbiol Rev 59:201–222Google Scholar
- Taylor RT, Jenkins WT (1966) Leucine aminotransferase. II. Purification and characterization J Biol Chem 241:4396–4405Google Scholar
- Tsuchida T, Yoshinaga F, Kubota K, Momose H, Okumura S (1975) Cultural conditions for l-leucine production by strain no 218, a mutant of Brevibacterium-Lactofermentum 2256. Agric Biol Chem 39:1149–1153Google Scholar
- Wang BW, Shi AQ, Tu R, Zhang XL, Wang QH, Bai FW (2012) Branched-Chain Higher Alcohols. In: Bai FW, Liu CG, Huang H, Tsao GT (eds) Biotechnology in China Iii: Biofuels and Bioenergy, vol 128. Adv Biochem Eng Biotechnol. https://doi.org/10.1007/10_2011_121
- Yu RYYM, Chen LR (2005) The principle and method of biochemistry experiment. Peking University Press, BeijingGoogle Scholar
- Zhu GJ (1994) Industrial microbiology experiment technical handbook. China light Industry Press, BeijingGoogle Scholar