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PLA (3-D-phenyllactic acid) is an ideal antimicrobial and immune regulatory compound present in honey and fermented foods. Sporolactobacillus inulinus is regarded as a potent D-PLA producer that reduces phenylpyruvate (PPA) with D-lactate dehydrogenases. In this study, PLA was produced by whole-cell bioconversion of S. inulinus ATCC 15538. Three genes encoding D-lactate dehydrogenase (d-ldh1, d-ldh2, and d-ldh3) were cloned and expressed in Escherichia coli BL21 (DE3), and their biochemical and structural properties were characterized. Consequently, a high concentration of pure D-PLA (47 mM) was produced with a high conversion yield of 88%. Among the three enzymes, D-LDH1 was responsible for the efficient conversion of PPA to PLA with kinetic parameters of Km (0.36 mM), k cat (481.10 s -1 ), and k cat /Km (1336.39 mM -1 s -1 ). In silico structural analysis and site-directed mutagenesis revealed that the Ile307 in D-LDH1 is a key residue for excellent PPA reduction with low steric hindrance at the substrate entrance. This study highlights that S. inulinus ATCC 15538 is an excellent PLA producer, equipped with a highly specific and efficient D-LDH1 enzyme.

Titel:	Biological characterization of D-lactate dehydrogenase responsible for high-yield production of D-phenyllactic acid in Sporolactobacillus inulinus.
Autor/in / Beteiligte Person:	Cheng, YY ; Park, TH ; Seong, H ; Kim, TJ ; Han, NS
Link:	Zum Volltext
Zeitschrift:	Microbial biotechnology, Jg. 15 (2022-11-01), Heft 11, S. 2717
Veröffentlichung:	Hoboken, NJ : Wiley-Blackwell ; <i>Original Publication</i>: Oxford : Blackwell, c2008-, 2022
Medientyp:	academicJournal
ISSN:	1751-7915 (electronic)
DOI:	10.1111/1751-7915.14125
Schlagwort:	Polyesters metabolism Lactate Dehydrogenases genetics Lactate Dehydrogenases metabolism Escherichia coli genetics Escherichia coli metabolism
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article; Research Support, Non-U.S. Gov't Language: English [Microb Biotechnol] 2022 Nov; Vol. 15 (11), pp. 2717-2729. <i>Date of Electronic Publication: </i>2022 Aug 03. MeSH Terms: Lactate Dehydrogenases* / genetics ; Lactate Dehydrogenases* / metabolism ; Escherichia coli* / genetics ; Escherichia coli* / metabolism ; Polyesters / metabolism References: BMC Microbiol. 2017 Dec 11;17(1):231. (PMID: 29228901) ; Appl Microbiol Biotechnol. 2017 May;101(9):3677-3687. (PMID: 28190098) ; Molecules. 2017 Nov 15;22(11):. (PMID: 29140277) ; J Microbiol Biotechnol. 2014 Dec 28;24(12):1664-72. (PMID: 25085568) ; Food Chem. 2015 Nov 1;186:265-70. (PMID: 25976820) ; Biotechnol Appl Biochem. 2017 Mar;64(2):279-289. (PMID: 26671214) ; Food Microbiol. 2020 Apr;86:103344. (PMID: 31703877) ; PLoS Genet. 2019 May 23;15(5):e1008145. (PMID: 31120900) ; Appl Microbiol Biotechnol. 2016 Sep;100(17):7471-8. (PMID: 27020295) ; PLoS One. 2011 Apr 20;6(4):e19030. (PMID: 21533054) ; J Appl Microbiol. 2021 May;130(5):1412-1424. (PMID: 32955761) ; Biosci Biotechnol Biochem. 2012;76(4):853-5. (PMID: 22484960) ; J Bacteriol. 2003 Aug;185(16):5023-6. (PMID: 12897026) ; J Food Sci Technol. 2019 May;56(5):2605-2610. (PMID: 31168142) ; Appl Microbiol Biotechnol. 2012 Sep;95(5):1155-63. (PMID: 22782253) ; J Food Sci. 2017 Oct;82(10):2269-2275. (PMID: 28881036) ; Lett Appl Microbiol. 2014 Oct;59(4):404-11. (PMID: 24888766) ; Biochem Biophys Res Commun. 1993 Apr 15;192(1):182-8. (PMID: 8476420) ; 3 Biotech. 2020 Mar;10(3):101. (PMID: 32099742) ; Int J Biol Macromol. 2021 May 15;179:71-79. (PMID: 33631263) ; Bioresour Technol. 2009 Nov;100(21):5226-9. (PMID: 19501505) ; J Dairy Sci. 2015 Feb;98(2):813-7. (PMID: 25434344) ; Int J Biol Macromol. 2015 Mar;74:608-19. (PMID: 25542170) ; Microb Biotechnol. 2022 Nov;15(11):2717-2729. (PMID: 35921426) ; Curr Microbiol. 2007 May;54(5):396-404. (PMID: 17387552) ; Enzyme Microb Technol. 2019 Jun;125:37-44. (PMID: 30885323) ; Front Bioeng Biotechnol. 2021 Feb 05;9:612105. (PMID: 33634085) ; Front Public Health. 2021 Jan 08;8:587298. (PMID: 33490017) ; Microbiologyopen. 2019 May;8(5):e00704. (PMID: 30066438) ; Appl Microbiol Biotechnol. 2017 Nov;101(22):8223-8236. (PMID: 28983655) ; Bioresour Technol. 2017 Dec;245(Pt B):1664-1673. (PMID: 28688739) ; Int J Syst Evol Microbiol. 2017 Jul;67(7):2363-2369. (PMID: 28699867) ; J Appl Microbiol. 2020 Aug;129(2):335-344. (PMID: 32009287) ; Biotechnol J. 2019 May;14(5):e1800656. (PMID: 30810274) ; Sci Rep. 2016 Apr 25;6:24887. (PMID: 27109778) ; Microorganisms. 2021 Nov 21;9(11):. (PMID: 34835522) ; J Biosci Bioeng. 2016 May;121(5):479-83. (PMID: 26472127) ; Int J Syst Evol Microbiol. 2008 Oct;58(Pt 10):2316-20. (PMID: 18842848) ; Biotechnol Lett. 2007 Apr;29(4):593-7. (PMID: 17333468) ; Biotechnol Lett. 2012 May;34(5):907-11. (PMID: 22261863) ; Food Funct. 2021 Jun 21;12(12):5591-5606. (PMID: 34017972) ; J Food Sci. 2011 Aug;76(6):M305-11. (PMID: 21729073) ; 3 Biotech. 2019 Mar;9(3):89. (PMID: 30800600) ; Molecules. 2015 Aug 24;20(8):15359-73. (PMID: 26305241) Substance Nomenclature: EC 1.1.1.28 (D-lactate dehydrogenase) ; 156-05-8 (3-phenyllactic acid) ; EC 1.1.- (Lactate Dehydrogenases) ; X7CO62M413 (phenylpyruvic acid) ; 0 (Polyesters) SCR Organism: Sporolactobacillus inulinus Entry Date(s): Date Created: 20220803 Date Completed: 20221101 Latest Revision: 20221130 Update Code: 20231215 PubMed Central ID: PMC9618312

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Biological characterization of D-lactate dehydrogenase responsible for high-yield production of D-phenyllactic acid in Sporolactobacillus inulinus.