Cloning, Characterization, and Expression of cDNA Encoding a Lipase from<i>Kurtzmanomyces</i>sp. I-11

Kakugawa, Koji; Shobayashi, Megumi; Сузукі, Осаму; Miyakawa, Tokichi

doi:10.1271/bbb.66.1328

Cited by 12 publications

(6 citation statements)

References 33 publications

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“…Therefore the discussion concerning the primary structure of these two lipases is described in the following report. 21) Both Candida antarctica and Kurtzmanomyces sp. I-11 produce high levels of the glycolipid type biosurfactants MEL in the culture broth.…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of a Lipase from the Glycolipid-Producing YeastKurtzmanomycessp. I-11

Kakugawa

Shobayashi

Сузукі

et al. 2002

Bioscience, Biotechnology, and Biochemistry

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An extracellular lipase produced by the glycolipidproducing yeast Kurtzmanomyces sp. I-11 was puriˆed by ammonium sulfate precipitation and column chromatographies on DEAE-Sephadex A-25, SP-Sephadex C-50, and Sephadex G-100. Based on the analysis of the puriˆed lipase on sodium dodecyl sulfatepolyacrylamide gel electrophoresis, the puriˆed lipase was judged to be homogeneous and its molecular mass was estimated to be approximately 49 kDa. The optimum temperature for the activity was 759 C, and the activity was very stable at temperatures below 709 C. The active pH range of this lipase was 1.9-7.2, and the activity was stable at pH below 7.1. The lipase showed a preference for C18 acyl groups by measurements with pnitrophenyl esters and triglycerides as substrates. The lipase was very stable in the presence of various organic solvents at a concentration of 40z. Although the Nterminal sequence of the Kurtzmanomyces lipase was very similar to that of lipase A from Candida antarctica, the pH proˆles of the two lipases were signiˆcantly diŠerent.

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Section: Discussionmentioning

confidence: 99%

Purification and Characterization of a Lipase from the Glycolipid-Producing YeastKurtzmanomycessp. I-11

Kakugawa

Shobayashi

Сузукі

et al. 2002

Bioscience, Biotechnology, and Biochemistry

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“…All further protein sequences identified during the BLAST search displayed an identity of less than 35%, indicating that these enzymes belong to different lipase subfamilies. While lipK was already functionally expressed in Pichia pastoris and characterized (Kakugawa et al 2002a;Kakugawa et al 2002b), the three sequences from the smut fungi U. maydis and S. reilianum are annotated as hypothetical lipases. With exception of the U. maydis sequence UM03410, all of the CAL-A-type lipases were predicted to be secretory proteins, which contain a cleavable, N-terminal signal sequence (Table 1).…”

Section: Structure-and Sequence-based Homology Searches On Cal-a-typementioning

confidence: 99%

The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity

Brundiek

Saß

Evitt

et al. 2012

Appl Microbiol Biotechnol

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The Ustilago maydis lipase UM03410 belongs to the mostly unexplored Candida antarctica lipase (CAL-A) subfamily. The two lipases with [corrected] the highest identity are a lipase from Sporisorium reilianum and the prototypic CAL-A. In contrast to the other CAL-A-type lipases, this hypothetical U. maydis lipase is annotated to possess a prolonged N-terminus of unknown function. Here, we show for the first time the recombinant expression of two versions of lipase UM03410: the full-length form (lipUMf) and an Nterminally truncated form (lipUMs). For comparison to the prototype, the expression of recombinant CAL-A in E. coli was investigated. Although both forms of lipase UM03410 could be expressed functionally in E. coli, the N-terminally truncated form (lipUMs) demonstrated significantly higher activities towards p-nitrophenyl esters. The functional expression of the N-terminally truncated lipase was further optimized by the appropriate choice of the E. coli strain, lowering the cultivation temperature to 20 °C and enrichment of the cultivation medium with glucose. Primary characteristics of the recombinant lipase are its pH optimum in the range of 6.5-7.0 and its temperature optimum at 55 °C. As is typical for lipases, lipUM03410 shows preference for long chain fatty acid esters with myristic acid ester (C14:0 ester) being the most preferred one.More importantly, lipUMs exhibits an inherent preference for C18:1Δ9 trans and C18:1Δ11 trans-fatty acid esters similar to CAL-A. Therefore, the short form of this U. maydis lipase is the only other currently known lipase with a distinct trans-fatty acid selectivity.

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“…Lipases from C. parapsilosis (Neugnot et al , 2002), Kurtzmanomyces sp. I‐11 (Kakugawa et al , 2002), and C. antarctica were shown to have 33%, 32%, and 33% identities, respectively, and putative lipases or unnamed proteins from Debaryomyces hansenii , Cordyceps bassiana , and Zygoascus hellenicus were shown to have 32%, 31%, and 31% identities, respectively. We speculate that M. pachydermatis , the nonlipophilic species, and other lipophilic Malassezia species produce lipases with different homology patterns due to the difference in lipid usage and pathogenesis.…”

Section: Resultsmentioning

confidence: 99%

Isolation, characterization and molecular cloning of a lipolytic enzyme secreted fromMalassezia pachydermatis

Shibata

Okanuma

Hirai

et al. 2006

FEMS Microbiology Letters

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Lipophilic Malassezia species may induce catheter-associated sepsis in premature neonates and immunocompromised patients receiving parenteral lipid emulsions. To assess the participation of lipolytic enzymes in the pathogenesis of this yeast, we cloned a gene encoding the enzyme. A lipolytic enzyme in the culture supernatant of Malassezia pachydermatis was purified 210-fold to homogeneity. The enzyme showed high esterase activity toward p-nitrophenyl octanoate. The cDNA encoding the enzyme was cloned using a degenerate oligonucleotide primer constructed from the N-terminal amino acid sequence. The cDNA consisted of 1582 bp, including an open reading frame encoding 470 amino acids. The first 19 amino acids and the following 13 amino-acid sequence were predicted to be the signal peptides for secretion and prosequence, respectively. The predicted molecular mass of the 438-amino acid mature protein was 48 kDa. Analysis of the deduced amino acid sequence revealed that it contains the consensus motif (Gly-X-Ser-X-Gly), which is conserved among lipolytic enzymes. Homology investigations showed that the enzyme has similarities principally with 11 lipases produced by Candida albicans (29-34% identity) and some other yeast lipases.

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Cloning, Characterization, and Expression of cDNA Encoding a Lipase fromKurtzmanomycessp. I-11

Cited by 12 publications

References 33 publications

Purification and Characterization of a Lipase from the Glycolipid-Producing YeastKurtzmanomycessp. I-11

Purification and Characterization of a Lipase from the Glycolipid-Producing YeastKurtzmanomycessp. I-11

The short form of the recombinant CAL-A-type lipase UM03410 from the smut fungus Ustilago maydis exhibits an inherent trans-fatty acid selectivity

Isolation, characterization and molecular cloning of a lipolytic enzyme secreted fromMalassezia pachydermatis

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