Hyperexpression and Analysis ofchoBEncoding Cholesterol Oxidase ofBrevibacterium sterolicuminEscherichia coliandStreptomyces lividans.

Ohta, Toshio; Fujishiro, Kinya; Yamaguchi, Kazuo; Uwajima, Takayuki; Aisaka, Kazuo; Hasegawa, Mamoru

doi:10.1271/bbb.56.1786

Cited by 24 publications

(14 citation statements)

References 11 publications

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“…This compound was also detected among the metabolites of cholesterol, or 4-cholesten-3-one by extra-or intracellular ChO from recombinant strains Streptomyces lividans and Escherichia coli with cloned DNA-fragment from Brevibacterium sterolicum. As established by genetic analysis, the enzyme responsible for the formation of 6-hydroxy derivative is ChO [9]. Similar results were described for commercial ChO (EC 1.1.3.6.)…”

Section: Introductionsupporting

confidence: 76%

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Extracellular 3β-hydroxysteroid oxidase of Mycobacterium vaccae VKM Ac-1815D

Nikolayeva

Egorova

Dovbnya

et al. 2004

The Journal of Steroid Biochemistry and Molecular Biology

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Section: Introductionsupporting

confidence: 76%

“…Enzyme molecular weight (62 ± 4 kDa), properties and multifunctional action corresponded to ChO from the relative organisms, rather than short-chain dehydrogenases [9][10][11].…”

Section: Discussionmentioning

confidence: 99%

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Extracellular 3β-hydroxysteroid oxidase of Mycobacterium vaccae VKM Ac-1815D

Nikolayeva

Egorova

Dovbnya

et al. 2004

The Journal of Steroid Biochemistry and Molecular Biology

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“…[12,16] and Arthrobacter simplex [17], choB from Brevibacterium sterolicum [18,19], choL from Streptomyces virginiae [20]. The expression of choB from B. sterolicum in Streptomyces lividans resulted in 85 times increased secretion of the active enzyme as compared with a wild-type strain [21]. The structural gene for ChoA A of A. simplex fusant F2 have been cloned and expressed in Escherichia coli and Pichia pastoris.…”

Section: Introductionmentioning

confidence: 99%

Cholesterol oxidase ChoD is not a critical enzyme accounting for oxidation of sterols to 3-keto-4-ene steroids in fast-growing Mycobacterium sp. VKM Ac-1815D

Ивашина

Nikolayeva

Dovbnya

et al. 2012

The Journal of Steroid Biochemistry and Molecular Biology

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“…sterolicum ATCC 21387 was the only presumptive non-R. equi strain included in our study which yielded a 0.95-kb PCR product with the COX primers. ATCC 21387 is a wellknown strain used for the industrial production of cholesterol oxidase and the extensive genetic and biochemical characterization of this enzyme (20,23,24). Indeed, the three-dimensional crystal structure of the cholesterol oxidase protein was determined by using the purified enzyme from B. sterolicum ATCC 21387 (15).…”

Section: Discussionmentioning

confidence: 99%

Rapid Identification ofRhodococcus equiby a PCR Assay Targeting thechoEGene

et al. 2003

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The actinomycete Rhodococcus equi is an important pathogen of horses and an emerging opportunistic pathogen of humans. Identification of R. equi by classical bacteriological techniques is sometimes difficult, and misclassification of an isolate is not uncommon. We report here on a specific PCR assay for the rapid and reliable identification of R. equi. It is based on the amplification of a fragment of the choE gene encoding cholesterol oxidase. The choE-based PCR was assessed by using a panel of strains comprising 132 isolates from different sources and of different geographical origins, all initially identified biochemically as R. equi, and 30 isolates of representative non-R. equi actinomycete species, including cholesterol oxidase producers. The expected 959-bp amplicon was observed only with R. equi isolates, as confirmed by sequencing of a variable region of the 16S RNA gene from a random sample of 20 PCR-positive isolates. All R. equi isolates gave a positive choE-based PCR result, which correlated with a high degree of conservation of the choE gene. Three of the 132 strains originally identified as R. equi were negative for the choE gene, and subsequent analysis of their 16S RNA gene sequences confirmed that they belonged to other bacterial species (Dietzia maris, Mycobacterium peregrinum, and Staphylococcus epidermidis). All non-R. equi isolates were negative by the choE-based PCR. ATCC 21387, the only known isolate of Brevibacterium sterolicum, gave a 959-bp amplicon whose DNA sequence was virtually identical to that of R. equi choE. Comparison of the 16S RNA genes indicated that ATCC 21387 should be considered an R. equi isolate.The nocardioform actinomycete Rhodococcus equi is a primary pathogen of horses. In foals, R. equi causes severe pyogranulomatous pneumonia, often accompanied by ulcerative enteritis and mesenteric lymphadenitis (29). In recent years, R. equi has emerged as an important opportunistic pathogen in humans, causing potentially life-threatening infections in severely immunocompromised people, in particular, human immunodeficiency virus-infected patients (43). In humans, R. equi causes a lung disease reminiscent of pulmonary tuberculosis. R. equi can also infect cattle, in which it has been associated with ulcerative lymphangitis (34), and it has also been recovered from inflamed tonsils and the cervical lymph nodes of pigs (21, 34). The natural habitat of R. equi is the soil, especially that enriched with fecal material from domestic and wild animals (37).R. equi infections are diagnosed by culturing and subsequent phenotypic analysis of the isolates by means of classical morphological and biochemical tests (9). However, the colony characteristics, cellular morphology, and reaction to acid-fast staining differ between R. equi isolates (32). Although the API Coryne multisubstrate identification system (bio-Merieux), a commercial kit widely used in clinical microbiology laboratories, includes R. equi in its database, its reliability for the biochemical identification of rhodococcal iso...

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Hyperexpression and Analysis ofchoBEncoding Cholesterol Oxidase ofBrevibacterium sterolicuminEscherichia coliandStreptomyces lividans.

Cited by 24 publications

References 11 publications

Extracellular 3β-hydroxysteroid oxidase of Mycobacterium vaccae VKM Ac-1815D

Extracellular 3β-hydroxysteroid oxidase of Mycobacterium vaccae VKM Ac-1815D

Cholesterol oxidase ChoD is not a critical enzyme accounting for oxidation of sterols to 3-keto-4-ene steroids in fast-growing Mycobacterium sp. VKM Ac-1815D

Rapid Identification ofRhodococcus equiby a PCR Assay Targeting thechoEGene

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