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44. Production of 5‐aminolevulinic acid from hydrolysates of cassava residue and fish waste by engineered Bacillus cereusPT1.

Author

Luo, Ying, Su, Anping, Yang, Jinshui, Yu, Qijun, Wang, Entao, and Yuan, Hongli

Subjects
*FISH waste, *BACILLUS (Bacteria), *CASSAVA, *AGRICULTURE, *BACILLUS cereus, *CUCUMBERS, *TRANSFER RNA, *ERGOT alkaloids
Abstract

The economical production of 5‐aminolevulinic acid (ALA) has recently received increasing attention for its extensive use in agriculture. In this study, a strain of Bacillus cereus PT1 could initially produce ALA at a titre of 251.72 mg/L by using a hydrolysate mixture of low‐cost cassava residue and fish waste. The integration of endogenous hemA encoding glutamyl‐tRNA reductase led to a 39.30% increase in ALA production. Moreover, improving cell permeability by deletion of the LytR‐CpsA‐Psr (LCP) family gene tagU led to a further increase of 59.73% in ALA production. Finally, the engineered strain B. cereus PT1‐hemA‐ΔtagU produced 2.62 g/L of ALA from the previously mentioned hydrolysate mixture in a 7‐L bioreactor. In a pot experiment, foliar spray of the ALA produced by B. cereus PT1‐hemA‐ΔtagU from the hydrolysates increased salt tolerance of cucumber by improving chlorophyll content and catalase activity, while decreasing malondialdehyde content. Overall, this study demonstrated an economic way to produce ALA using a microbial platform and evidenced the potential of ALA in agricultural application. [ABSTRACT FROM AUTHOR]

Published
2023
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50. Efficient biosynthesis of 5-aminolevulinic acid from glutamate viawhole-cell biocatalyst in immobilized engineered Escherichia coliElectronic supplementary information (ESI) available. See DOI: https://doi.org/10.1039/d2cy02172b

Author

Luo, Ying, Liu, Liang, Yang, Jinshui, Su, Anping, Yu, Qijun, Wang, Entao, and Yuan, Hongli

Abstract

5-Aminolevulinic acid (ALA), a natural plant growth regulator, herbicide, insecticide, and photodynamic therapy drug, has various applications in agriculture and medicine. Currently, ALA mainly relies on chemical synthesis; however, this method is unsustainable and cannot meet the increasing demand because of its low yields. In this study, through phylogenetic analysis and the statistic of gene numbers downstream to hemA, we selected 4 candidate hemAgenes with high potential for ALA production from genome sequences of 562 bacterial strains and found that hemAfrom Burkholderia caryophyllihad the best effect on ALA production. Next, we found the match of HemA and homologous glutamyl-tRNAGlu, and further properly overexpressing gltXand tRNAGluincreased glutamyl-tRNAGlurecognition by HemA. Moreover, through the rational combination of gene elements from B. caryophylli(hemA, tRNAGlu) and Salmonella typhimurium(gltX, hemL), optimizing the gene expression model, as well as E. colihosts, the efficiency of ALA synthesis through the C5 pathway was significantly enhanced in engineered E. coliBurAt-SalSL1 (T7). Finally, we developed a whole-cell bioconversion system to produce ALA for the first time and 20.89 g ALA was obtained from 42.42 g glutamate with a conversion of 63.51% after six catalytic recycles. Overall, our study provides an alternative route for the green production of ALA.

Published
2023
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