Purification and Characterization of a Novel Cholesterol Esterase from<i>Pseudomonas aeruginosa</i>, with Its Application to Cleaning Lipid-stained Contact Lenses

Sugihara, Akio; Shimada, Yuji; Nomura, Atsuo; Terai, Tadamasa; Imayasu, Masaki; Nagai, Yusuke; Nagao, Toshihiro; Watanabe, Yomi; Tominaga, Yoshio

doi:10.1271/bbb.66.2347

Cited by 27 publications

(19 citation statements)

References 21 publications

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“…These data were similar to those of P. aeruginosa (CHE-A) 12 , Streptomyces sp. X9 35 , and S. avermitilis 30 .…”

Section: Discussionsupporting

confidence: 87%

“…According to PAS staining test, CHEPaRE24.3 carried a 60 kDa oligosaccharide, which is a similar finding to CHE in P. aeruginosa 12 . Referring to optimum pH and temperature data, the enzyme showed similar features to those of Streptomyces sp.…”

Section: Discussionsupporting

confidence: 72%

“…The activity was assayed in 100 mM sodium phosphate buffer (pH 7.0) at 4°C. The enzyme activities were compared among CHEPaRE24.3 (grey bars), CHE-A (horizontal striped bars) 12 , COE-311 (white bars) (Pseudomonas sp., strain COE-311 in Toyobo Enzyme Manual 12 ), CHE-1 (vertical striped bars) (Pseudomonas sp., strain CHE-1 in Amano Enzyme Manual 12 ), and P. fluorescens (black bars) 11 . The standard error was less than 5% when triplicate reactions were tested.…”

Section: Substrate Specificitymentioning

confidence: 99%

“…CHE has been extensively examined in a number of mammalian tissues such as pancreas 7 , small intestine 7 , brain 8 , liver 9 , and placenta 10 . The enzyme also has a widespread distribution in bacteria, fungi, and yeasts such as Pseudomonas fluorescens 11 , P. aeruginosa 12 , Fusarium oxysporum 13 , Candida cylindracea 14 , Saccharomyces cerevisiae 15 , Staphylococcus aureus 16 and Streptomyces lavendulae 17 . The practical use of these enzymes is limited to the measurement of total cholesterol in combination with cholesterol oxidase (EC 1.1.3.6) in blood.…”

Section: Introductionmentioning

confidence: 99%

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Khuchareontaworn¹,

Pakpitchareon²,

Areekit³

et al. 2010

ScienceAsia

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Cholesterol esterase (CHE; EC 3.1.1.13) plays an important role in the hydrolysis of cholesterol ester to cholesterol and free fatty acids. It is used for clinical detection of cholesterol. Herein, the CHE producing microorganisms were isolated from oil contaminated soil samples at 45°C by spreading the sample on nutritive agar. Pseudomonas aeruginosa strain RE 24.3 was selected since it produced the highest CHE activity. The purified enzyme, namely CHEPaRE24.3, had high activity between 30 and 45°C at the optimum pH of 7.0. The substrate specificity test revealed that CHEPaRE24.3 hydrolysed a variety of cholesterol esters. Inhibitor examination found that Hg 2+ , β-mercaptoethanol, and DTT could inhibit the enzyme activity at 99.38%, 100%, and 43.2%, respectively. It is remarkable that PMSF showed no effect on enzyme activity. The stability test indicated that the CHEPaRE24.3 enzyme remained at 70% activity after 28 days storage at 4°C, 29°C, or 37°C. The long-term storage stability and thermotolerance of CHEPaRE24.3 as well as its resistance to PMSF suggests that it could be applicable for use in a cholesterol biosensor.

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“…These data were similar to those of P. aeruginosa (CHE-A) 12 , Streptomyces sp. X9 35 , and S. avermitilis 30 .…”

Section: Discussionsupporting

confidence: 87%

Section: Discussionsupporting

confidence: 72%

Section: Substrate Specificitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Untitled

Khuchareontaworn¹,

Pakpitchareon²,

Areekit³

et al. 2010

ScienceAsia

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“…Furthermore, cell wall teichoic acids of S. aureus confer resistance to fatty acids from skin sebaceous glands [87] . At this time it is still speculative whether a cholesterol esterase produced by P. aeruginosa [88] may represent an additional virulence factor aiding in the inactivation of host-derived antimicrobial cholesteryl esters. Of interest is the recent finding of Cadieux et al [89] who identified a lipase in a hypervirulent community-associated methicillin-resistant S. aureus strain USA300 that hydrolyzes triglycerides and liberates the free fatty acid linoleic acid with growth inhibitory activity against S. aureus.…”

Section: Bacterial Mechanisms That Manipulate Host-derived Lipidsmentioning

confidence: 99%

Antimicrobial lipids: Emerging effector molecules of innate host defense

Porter

Daniel²,

Alvarez³

et al. 2015

WJI

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The antimicrobial properties of host-derived derived lipids have become increasingly recognized and evidence is mounting that antimicrobial lipids (AMLs), like antimicrobial peptides, are effector molecules of the innate immune system and are regulated by its conserved pathways. This review, with primary focus on the human body, provides some background on the biochemistry of lipids, summarizes their biological functions, expands on their antimicrobial properties and site-specific composition, presents modes of synergism with antimicrobial peptides, and highlights the more recent reports on the regulation of AML production as well as bacterial resistance mechanisms. Based on extant data a concept of innate epithelial defense is proposed where epithelial cells, in response to microbial products and proinflammatory cytokines and through activation of conserved innate signaling pathways, increase their lipid uptake and up-regulate transcription of enzymes involved in antimicrobial lipid biosynthesis, and induce transcription of antimicrobial peptides as well as cytokines and chemokines. The subsequently secreted antimicrobial peptides and lipids then attack and eliminate the invader, assisted by or in synergism with other antimicrobial molecules delivered by other defense cells that have been recruited to the site of infection, in most of the cases. This review invites

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Cloning and expression of a Melanocarpus albomyces steryl esterase gene in Pichia pastoris and Trichoderma reesei

Kontkanen

Reinikainen

Saloheimo

2006

Biotech & Bioengineering

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The ste1 gene encoding a steryl esterase was isolated from the thermophilic fungus Melanocarpus albomyces. The gene has one intron, and it encodes a protein consisting of 576 amino acids. The deduced amino acid sequence of the steryl esterase was shown to be related to lipases and other esterases such as carboxylesterases. Formation of mature protein requires post-translational removal of a putative 18-amino-acid signal sequence and a 13-residue propeptide at the N-terminus. The intronless version of the Melanocarpus albomyces ste1 gene was expressed in Pichia pastoris under the inducible AOX1 promoter. The production level was low, and a large proportion of the total activity yield was found to be present intracellularly. However, the fact that steryl esterase activity was produced by P. pastoris cells carrying the expression cassette confirmed that the correct gene had been cloned. The ste1 gene was subsequently expressed in T. reesei under the inducible cbh1 promoter, and a clearly higher production level was obtained. About 60% of the total activity was bound to the fungal mycelium or to solid components of the culture medium, or existed as aggregates. Triton X-100 was successfully used to recover this activity. The heterologous production system in T. reesei provides a means of producing M. albomyces steryl esterase STE1 reliably in large scale for future studies.

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Purification and Characterization of a Novel Cholesterol Esterase fromPseudomonas aeruginosa, with Its Application to Cleaning Lipid-stained Contact Lenses

Cited by 27 publications

References 21 publications

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Antimicrobial lipids: Emerging effector molecules of innate host defense

Cloning and expression of a Melanocarpus albomyces steryl esterase gene in Pichia pastoris and Trichoderma reesei

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