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Production of 12-hydroxy Dodecanoic Acid Methyl Ester Using a Signal Peptide Sequence-Optimized Transporter AlkL and a Novel Monooxygenase

Production of 12-hydroxy Dodecanoic Acid Methyl Ester Using a Signal Peptide Sequence-Optimized Transporter AlkL and a Novel Monooxygenase

Hee-Wang Yoo, Joonwon Kim, Mahesh D Patil, Beom Gi Park, Sung-Yeon Joo, Hyungdon Yun, Byung-Gee Kim

Bioresour Technol. 2019 Nov;291:121812.

PMID: 31376668

Abstract:

In this study, a signal peptide of AlkL was replaced with other signal peptides to improve the soluble expression and thereby facilitate the transport of dodecanoic acid methyl ester (DAME) substrate into the E. coli. Consequently, AlkL with signal peptide FadL (AlkLf) showed higher transport activity toward DAME. Furthermore, the promoter optimization for the efficient heterologous expression of the transporter AlkLf and alkane monooxygenase (AlkBGT) system was conducted and resulted in increased ω-oxygenation activity of AlkBGT system. Moreover, bioinformatic studies led to the identification of novel monooxygenase from Pseudomonas pelagia (Pel), which exhibited 20% higher activity towards DAME as substrate compared to AlkB. Finally, the construction of a chimeric transporter and the expression of newly identified monooxygenase enabled the production of 44.8 ± 7.5 mM of 12-hydroxy dodecanoic acid methyl ester (HADME) and 31.8 ± 1.7 mM of dodecanedioic acid monomethyl ester (DDAME) in a two-phase reaction system.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP96801397	12-(7-Nitrobenzofurazan-4-ylamino)dodecanoic acid		96801-39-7	Price

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