Purification and Characterization of Isocitrate Lyase from the Wood-Destroying Basidiomycete Fomitopsis palustris Grown on Glucose

Munir, Erman; Hattori, Takefumi; Shimada, Morimi

doi:10.1006/abbi.2002.2770

Cited by 12 publications

(10 citation statements)

References 45 publications

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“…The deduced polypeptide had a predicted molecular mass of 59 735 Da, which is similar to the estimated molecular mass for the purified FPICL1 (Munir et al, 2002). By analogy with acu-D, the ICLs of A. nidulans (Gainey et al, 1992), D111, D165, D167 and E194 are suggested to be a cluster for the Mg 2+ -binding site.…”

Section: Characterization Of Fpicl1supporting

confidence: 53%

“…Purified ICL protein (approx. 80 mg; Munir et al, 2002) was digested with lysylendopeptidase, and the resulting peptides were separated by HPLC on a reverse-phase C18 column. N-terminal amino acid sequences were determined by use of an ABI Precise 491 automated protein sequencer (Applied Biosystems).…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, we were motivated to investigate the intracellular localization of the GLOX cycle enzymes, and other enzymes involved in oxalate fermentation, in glucose-grown F. palustris. Previously, we purified and characterized ICL from F. palustris (Munir et al, 2002), which is termed FPICL1 in this study. We report here the characterization of FPICL1 cDNA encoding FPICL1, and peroxisomal localization of FPICL1 based on subcellular fractionation and immunocytochemical experiments using anti-FPICL1 antibody.…”

Section: Introductionmentioning

confidence: 99%

“…The immunoreactive proteins were detected by chemiluminescence with the ECL detection system (Amersham Biosciences), according to the manufacturer's instructions. Preparation of cell-free extracts from F. palustris, and purification of the native enzyme, were performed by the methods of Munir et al (2002).…”

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confidence: 99%

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Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose

et al. 2006

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This study investigated the subcellular localization of key enzymes of the glyoxylate cycle, i.e. isocitrate lyase (ICL; EC 4.1.3.1) and malate synthase (EC 2.3.3.9), that function constitutively in coordination with oxalate biosynthesis of glucose-grown Fomitopsis palustris. The ICL purified previously from F. palustris is termed FPICL1. Subcellular fractionation analysis of the cell homogenate by the sucrose density-gradient method showed that both key enzymes were present in peroxisomes, whereas acetyl-CoA synthase (EC 6.2.1.1) and oxalate-producing oxaloacetate acetylhydrolase (EC 3.7.1.1) were cytosolic. The peroxisomal localization of FPICL1 was further confirmed by electron microscopic and immunocytochemical analysis with anti-FPICL1 antibody. In addition, the peroxisomal target signal, composed of SKL at the C terminus of the cDNA encoding FPICL1, was found, which also suggests that FPICL1 is peroxisomal. Accordingly, it is postulated that transportation of succinate from peroxisomes to mitochondria, and vice versa, for the transportation of isocitrate or citrate, occurs in glucose-grown F. palustris for the constitutive metabolic coordination of the TCA and glyoxylate cycles with oxalate biosynthesis.

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Section: Characterization Of Fpicl1supporting

confidence: 53%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose

et al. 2006

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“…Moreover, we have suggested that F. palustris transports oxalate out of the cells to prevent possible inhibition of intracellular metabolic reactions. For example, oxalate might inhibit the activities of FPICL1 and malate synthase in peroxisomes of F. palustris (15,35), because oxalate competitively inhibits the activities of the two enzymes in vitro (30,31). In addition, oxalate produced from oxaloacetate by cytosolic OAAH in F. palustris (1,35) possibly shows product inhibition for OAAH activity because oxalate is a competitive inhibitor (K i ϭ 19 M) for OAAH from Botrytis cinerea (13).…”

Section: Discussionmentioning

confidence: 99%

Oxalate Efflux Transporter from the Brown Rot Fungus Fomitopsis palustris

Watanabe

Shitan

Suzuki

et al. 2010

Appl Environ Microbiol

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An oxalate-fermenting brown rot fungus, Fomitopsis palustris, secretes large amounts of oxalic acid during wood decay. Secretion of oxalic acid is indispensable for the degradation of wood cell walls, but almost nothing is known about the transport mechanism by which oxalic acid is secreted from F. palustris hyphal cells. We characterized the mechanism for oxalate transport using membrane vesicles of F. palustris. Oxalate transport in F. palustris was ATP dependent and was strongly inhibited by several inhibitors, such as valinomycin and NH 4 ؉ , suggesting the presence of a secondary oxalate transporter in this fungus. We then isolated a cDNA, FpOAR (Fomitopsis palustris oxalic acid resistance), from F. palustris by functional screening of yeast transformants with cDNAs grown on oxalic acid-containing plates. FpOAR is predicted to be a membrane protein that possesses six transmembrane domains but shows no similarity with known oxalate transporters. The yeast transformant possessing FpOAR (FpOAR-transformant) acquired resistance to oxalic acid and contained less oxalate than the control transformant. Biochemical analyses using membrane vesicles of the FpOAR-transformant showed that the oxalate transport property of FpOAR was consistent with that observed in membrane vesicles of F. palustris. The quantity of FpOAR transcripts was correlated with increasing oxalic acid accumulation in the culture medium and was induced when exogenous oxalate was added to the medium. These results strongly suggest that FpOAR plays an important role in wood decay by acting as a secondary transporter responsible for secretion of oxalate by F. palustris.

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Isocitric acid production from rapeseed oil by Yarrowia lipolytica yeast

Kamzolova

Dedyukhina

Samoilenko

et al. 2013

Appl Microbiol Biotechnol

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Production of D S-threo-isocitric acid (ICA) by yeast meets serious difficulties since it is accompanied by a simultaneous production of citric acid (CA) in significant amounts that reduces the yield of desired product. In order to develop an effective process of ICA production, 60 yeast strains of different genera (Candida, Pichia, Saccharomyces, Torulopsis, and Yarrowia) were tested for their ability to produce ICA from rapeseed oil; as a result, wild-type strain Yarrowia lipolytica VKM Y-2373 and its mutant Y. lipolytica 704-UV4-A/NG50 were selected as promising ICA producers. The effects of temperature, pH, aeration, and concentrations of rapeseed oil, iron, and itaconic acid on ICA production by selected strains were studied. Under optimal conditions (pH 6.0; aeration 50-55 %; rapeseed oil concentration of 20-60 gl(-1), iron ion concentration of 1.2 mg l(-1), and itaconic acid amount of 30 mM), selected strains of Y. lipolytica produced predominantly ICA with a low amount of a by-product, CA.

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Purification and Characterization of Isocitrate Lyase from the Wood-Destroying Basidiomycete Fomitopsis palustris Grown on Glucose

Cited by 12 publications

References 45 publications

Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose

Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose

Oxalate Efflux Transporter from the Brown Rot Fungus Fomitopsis palustris

Isocitric acid production from rapeseed oil by Yarrowia lipolytica yeast

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