Effects of Expression of hemA and hemB Genes on Production of Porphyrin in Propionibacterium freudenreichii

DOI: 10.1128/AEM.70.12.7561-7566.2004

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FIG. 1.
Overview of the biosynthesis of tetrapyrroles with gene products indicated. Dashed lines denote multistep pathways, and broad arrows indicate nonenzymic steps.
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FIG. 2.
Profiles after HPLC of porphyrins from (A) a standard mixture of porphyrins, (B) a sample extracted from a culture of P. freudenreichii IFO12426 that harbored pPK705, and (C) a sample extracted from a culture of P. freudenreichii IFO12426 that harbored pKHEM04. Peaks: 1, UP I; 2, UP III; 3, various porphyrin derivatives; 4, CP III; 5, PP. Conditions for HPLC are described in the text.
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FIG. 3.
Time course of the production of porphyrins by P. freudenreichii IFO 12426 that harbored various expression vectors. (A) Production of UP I; (B) production of UP III; (C) production of CP. Open circles, pPK705; open triangles, pKHEM04; open squares, pKHEM05; open diamonds, pKHEM06. Each point is the mean of three experiments ± standard deviation.

TABLE 1.

Activities of ALA synthase and PBG synthase and production of ALA and PBG by P. freudenreichii harboring various expression vectors^a

Plasmid	Genotype	ALA synthase activity (μmol/min/mg of protein)^b	Production of ALA (μM)^c	PBG synthase activity (μmol/min/mg of protein)^b	Production of PBG (μM)^c
pPK705		0.26	35	0.15	2
pKHEM01	hemA⁺	0.71	62	0.20	12
pKHEM02	hemA⁺ hemB⁺	0.88	99	0.45	34
pKHEM03	hemB⁺	0.28	27	0.63	5
pKHEM04	hemA⁺	1.04	160	0.18	55
pKHEM05	hemA⁺ hemB⁺	1.19	159	0.56	72
pKHEM06	hemB⁺	0.26	35	0.74	4

↵ a Enzymatic activities and products were quantitated in triplicate under the conditions described in the text, and deviators were within 15%.
↵ b Cells were grown in GYT medium for 48 h.
↵ c Cells were grown in GYT medium for 72 h.