Optimization of a Two-Species Microbial Consortium for Improved Mcl-PHA Production From Glucose-Xylose Mixtures

Overview

Journal Front Bioeng Biotechnol

Date 2022 Jan 27

PMID 35083203

Authors

Yinzhuang Zhu

Mingmei Ai

Xiaoqiang Jia

Affiliations

Soon will be listed here.

Abstract

Polyhydroxyalkanoates (PHAs) have attracted much attention as a good substitute for petroleum-based plastics, especially mcl-PHA due to their superior physical and mechanical properties with broader applications. Artificial microbial consortia can solve the problems of low metabolic capacity of single engineered strains and low conversion efficiency of natural consortia while expanding the scope of substrate utilization. Therefore, the use of artificial microbial consortia is considered a promising method for the production of mcl-PHA. In this work, we designed and constructed a microbial consortium composed of engineered MG1655 and KT2440 based on the "nutrition supply-detoxification" concept, which improved mcl-PHA production from glucose-xylose mixtures. An engineered that preferentially uses xylose was engineered with an enhanced ability to secrete acetic acid and free fatty acids (FFAs), producing 6.44 g/L acetic acid and 2.51 g/L FFAs with 20 g/L xylose as substrate. The mcl-PHA producing strain of in the microbial consortium has been engineered to enhance its ability to convert acetic acid and FFAs into mcl-PHA, producing 0.75 g/L mcl-PHA with mixed substrates consisting of glucose, acetic acid, and octanoate, while also reducing the growth inhibition of by acetic acid. The further developed artificial microbial consortium finally produced 1.32 g/L of mcl-PHA from 20 g/L of a glucose-xylose mixture (1:1) after substrate competition control and process optimization. The substrate utilization and product synthesis functions were successfully divided into the two strains in the constructed artificial microbial consortium, and a mutually beneficial symbiosis of "nutrition supply-detoxification" with a relatively high mcl-PHA titer was achieved, enabling the efficient accumulation of mcl-PHA. The consortium developed in this study is a potential platform for mcl-PHA production from lignocellulosic biomass.

Citing Articles

Modification of Glucose Metabolic Pathway to Enhance Polyhydroxyalkanoate Synthesis in .

Dong Y, Zhai K, Li Y, Lv Z, Zhao M, Gan T Curr Issues Mol Biol. 2024; 46(11):12784-12799.

PMID: 39590355 PMC: 11592762. DOI: 10.3390/cimb46110761.

Bio-conversion of organic wastes towards polyhydroxyalkanoates.

Kuang Z, Yang H, Shen S, Lin Y, Sun S, Neureiter M Biotechnol Notes. 2024; 4:118-126.

PMID: 39416913 PMC: 11446391. DOI: 10.1016/j.biotno.2023.11.006.

Microbial Biosynthesis of Medium-Chain-Length Polyhydroxyalkanoate (mcl-PHA) from Waste Cooking Oil.

Elazzazy A, Ali Abd K, Bataweel N, Mahmoud M, Baghdadi A Polymers (Basel). 2024; 16(15).

PMID: 39125176 PMC: 11314287. DOI: 10.3390/polym16152150.

Medium Chain Length Polyhydroxyalkanoate Production by Engineered Pseudomonas gessardii Using Acetate-formate as Carbon Sources.

Kim W, Kim S, Seo H, Yang Y, Lee J, Hur M J Microbiol. 2024; 62(7):569-579.

PMID: 38700774 DOI: 10.1007/s12275-024-00136-x.

Efficient Bioremediation of Petroleum-Contaminated Soil by Immobilized Bacterial Agent of W33.

Yang Y, Zhang W, Zhang Z, Yang T, Xu Z, Zhang C Bioengineering (Basel). 2023; 10(5).

PMID: 37237630 PMC: 10215891. DOI: 10.3390/bioengineering10050561.

References

Anburajan P, Kumar A, Sabapathy P, Kim G, Cayetano R, Yoon J . Polyhydroxy butyrate production by Acinetobacter junii BP25, Aeromonas hydrophila ATCC 7966, and their co-culture using a feast and famine strategy. Bioresour Technol. 2019; 293:122062. DOI: 10.1016/j.biortech.2019.122062. View

Cavalheiro J, Pollet E, Diogo H, Cesario M, Averous L, de Almeida M . On the heterogeneous composition of bacterial polyhydroxyalkanoate terpolymers. Bioresour Technol. 2013; 147:434-441. DOI: 10.1016/j.biortech.2013.08.009. View

Wang H, Zhou X, Liu Q, Chen G . Biosynthesis of polyhydroxyalkanoate homopolymers by Pseudomonas putida. Appl Microbiol Biotechnol. 2010; 89(5):1497-507. DOI: 10.1007/s00253-010-2964-x. View

Tortajada M, da Silva L, Auxiliadora Prieto M . Second-generation functionalized medium-chain-length polyhydroxyalkanoates: the gateway to high-value bioplastic applications. Int Microbiol. 2013; 16(1):1-15. DOI: 10.2436/20.1501.01.175. View

Wang Q, Nomura C . Monitoring differences in gene expression levels and polyhydroxyalkanoate (PHA) production in Pseudomonas putida KT2440 grown on different carbon sources. J Biosci Bioeng. 2010; 110(6):653-9. DOI: 10.1016/j.jbiosc.2010.08.001. View

Liu X, Li X, Jiang J, Liu Z, Qiao B, Li F . Convergent engineering of syntrophic Escherichia coli coculture for efficient production of glycosides. Metab Eng. 2018; 47:243-253. DOI: 10.1016/j.ymben.2018.03.016. View

Lowe H, Hobmeier K, Moos M, Kremling A, Pfluger-Grau K . Photoautotrophic production of polyhydroxyalkanoates in a synthetic mixed culture of and . Biotechnol Biofuels. 2017; 10:190. PMC: 5517840. DOI: 10.1186/s13068-017-0875-0. View

Tobin K, OLeary N, Dobson A, OConnor K . Effect of heterologous expression of phaG [(R)-3-hydroxyacyl-ACP-CoA transferase] on polyhydroxyalkanoate accumulation from the aromatic hydrocarbon phenylacetic acid in Pseudomonas species. FEMS Microbiol Lett. 2007; 268(1):9-15. DOI: 10.1111/j.1574-6968.2006.00607.x. View

Yang S, Li S, Jia X . Production of medium chain length polyhydroxyalkanoate from acetate by engineered Pseudomonas putida KT2440. J Ind Microbiol Biotechnol. 2019; 46(6):793-800. DOI: 10.1007/s10295-019-02159-5. View

10.

Salas J, Ohlrogge J . Characterization of substrate specificity of plant FatA and FatB acyl-ACP thioesterases. Arch Biochem Biophys. 2002; 403(1):25-34. DOI: 10.1016/S0003-9861(02)00017-6. View

11.

Lee S . Bacterial polyhydroxyalkanoates. Biotechnol Bioeng. 1996; 49(1):1-14. DOI: 10.1002/(SICI)1097-0290(19960105)49:1<1::AID-BIT1>3.0.CO;2-P. View

12.

Zhao K, Deng Y, Chen J, Chen G . Polyhydroxyalkanoate (PHA) scaffolds with good mechanical properties and biocompatibility. Biomaterials. 2002; 24(6):1041-5. DOI: 10.1016/s0142-9612(02)00426-x. View

13.

Morgan-Sagastume F, Hjort M, Cirne D, Gerardin F, Lacroix S, Gaval G . Integrated production of polyhydroxyalkanoates (PHAs) with municipal wastewater and sludge treatment at pilot scale. Bioresour Technol. 2015; 181:78-89. DOI: 10.1016/j.biortech.2015.01.046. View

14.

Eiteman M, Altman E . Overcoming acetate in Escherichia coli recombinant protein fermentations. Trends Biotechnol. 2006; 24(11):530-6. DOI: 10.1016/j.tibtech.2006.09.001. View

15.

Zhang X, Li M, Agrawal A, San K . Efficient free fatty acid production in Escherichia coli using plant acyl-ACP thioesterases. Metab Eng. 2011; 13(6):713-22. DOI: 10.1016/j.ymben.2011.09.007. View

16.

Liang Y, Zhao W, Chen G . Study on the biocompatibility of novel terpolyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate). J Biomed Mater Res A. 2008; 87(2):441-9. DOI: 10.1002/jbm.a.31801. View

17.

Rodrigues P, Assis D, Druzian J . Simultaneous production of polyhydroxyalkanoate and xanthan gum: From axenic to mixed cultivation. Bioresour Technol. 2019; 283:332-339. DOI: 10.1016/j.biortech.2019.03.095. View

18.

Zhang H, Stephanopoulos G . Co-culture engineering for microbial biosynthesis of 3-amino-benzoic acid in Escherichia coli. Biotechnol J. 2016; 11(7):981-7. DOI: 10.1002/biot.201600013. View

19.

Jones J, Wang X . Use of bacterial co-cultures for the efficient production of chemicals. Curr Opin Biotechnol. 2017; 53:33-38. DOI: 10.1016/j.copbio.2017.11.012. View

20.

Chung A, Jin H, Huang L, Ye H, Chen J, Wu Q . Biosynthesis and characterization of poly(3-hydroxydodecanoate) by β-oxidation inhibited mutant of Pseudomonas entomophila L48. Biomacromolecules. 2011; 12(10):3559-66. DOI: 10.1021/bm200770m. View