Park Si Jae, Oh Young Hoon, Noh Won, Kim Hye Young, Shin Jae Ho, Lee Eun Gyo, Lee Seungwoon, David Yokimiko, Baylon Mary Grace, Song Bong Keun, Jegal Jonggeon, Lee Sang Yup, Lee Seung Hwan
Department of Environmental Engineering and Energy, Myongji University, Cheoin-gu, Yongin, Gyeonggido, Republic of Korea.
Biotechnol J. 2014 Oct;9(10):1322-8. doi: 10.1002/biot.201400156. Epub 2014 Sep 8.
L-Lysine is a potential feedstock for the production of bio-based precursors for engineering plastics. In this study, we developed a microbial process for high-level conversion of L-lysine into 5-aminovalerate (5AVA) that can be used as a monomer in nylon 6,5 synthesis. Recombinant Escherichia coli WL3110 strain expressing Pseudomonas putida delta-aminovaleramidase (DavA) and lysine 2-monooxygenase (DavB) was grown to high density in fed-batch culture and used as a whole cell catalyst. High-density E. coli WL3110 expressing DavAB, grown to an optical density at 600 nm (OD600 ) of 30, yielded 36.51 g/L 5AVA from 60 g/L L-lysine in 24 h. Doubling the cell density of E. coli WL3110 improved the conversion yield to 47.96 g/L 5AVA from 60 g/L of L-lysine in 24 h. 5AVA production was further improved by doubling the L-lysine concentration from 60 to 120 g/L. The highest 5AVA titer (90.59 g/L; molar yield 0.942) was obtained from 120 g/L L-lysine by E. coli WL3110 cells grown to OD600 of 60. Finally, nylon 6,5 was synthesized by bulk polymerization of ϵ-caprolactam and δ-valerolactam prepared from microbially synthesized 5AVA. The hybrid system demonstrated here has promising possibilities for application in the development of industrial bio-nylon production processes.
L-赖氨酸是生产工程塑料生物基前体的潜在原料。在本研究中,我们开发了一种微生物工艺,可将L-赖氨酸高效转化为5-氨基戊酸(5AVA),后者可用作尼龙6,5合成中的单体。表达恶臭假单胞菌δ-氨基戊酰胺酶(DavA)和赖氨酸2-单加氧酶(DavB)的重组大肠杆菌WL3110菌株在补料分批培养中生长至高密度,并用作全细胞催化剂。表达DavAB的高密度大肠杆菌WL3110在600nm处的光密度(OD600)达到30,在24小时内从60g/L的L-赖氨酸中产生了36.51g/L的5AVA。将大肠杆菌WL3110的细胞密度加倍,可使24小时内从60g/L的L-赖氨酸中的转化产率提高到47.96g/L的5AVA。将L-赖氨酸浓度从60g/L加倍至120g/L,进一步提高了5AVA的产量。通过将大肠杆菌WL3110细胞生长至OD600为60,从120g/L的L-赖氨酸中获得了最高的5AVA滴度(90.59g/L;摩尔产率0.942)。最后,通过由微生物合成的5AVA制备的ε-己内酰胺和δ-戊内酯的本体聚合反应合成了尼龙6,5。本文展示的混合系统在工业生物尼龙生产工艺开发中的应用具有广阔前景。