Factors Affecting Production of Staphylococcal Lipase

Vadehra, D. V.; Harmon, L. G.

doi:10.1111/j.1365-2672.1969.tb00958.x

Cited by 35 publications

(17 citation statements)

References 5 publications

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“…The strategy used for nucleotide sequencing is shown in Fig. 2 (28), it is likely that the codon at position 706 is the actual initiation codon. Furthermore, it is preceded 4 bp upstream by a potential Shine-Dalgarno sequence (GAGGTGAT) which matches exactly with the 3' terminus of E. coli 16S rRNA (23,25).…”

Section: Resultsmentioning

confidence: 99%

Lysogenic conversion of staphylococcal lipase is caused by insertion of the bacteriophage L54a genome into the lipase structural gene

Lee¹,

Iandolo²

1986

J Bacteriol

166

103

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Staphylococcus aureus PS54 manifests no lipase (geh) activity. This is due to the insertion of bacteriophage L54a DNA into the geh structural gene. The nucleotide sequence of this 2,968-base-pair DNA fragment was determined. Lipase deduced from the nucleotide sequence is a polypeptide of 690 amino acids which extends from nucleotide 706 to 2776.Many strains of staphylococci produce a true lipase or glycerol ester hydrolase (EC 3.1.1.3). The activity of the staphylococcal lipase gene is negatively regulated by bacteriophage lysogenization, also known as lysogenic conversion (3, 21 385(1 x SSC is 0.15 M NaCI and 15 mM sodium citrate [pH 7.0]) containing 0.1% sodium dodecyl sulfate. The membrane was dried at 80°C for 10 min and then autoradiographed as we described previously (4).DNA sequence analysis. Various restriction endonuclease fragments of the geh element were cloned into the M13 bacteriophage derivatives mpl8 or mpl9 (33) and propagated in Escherichia coli JM103. DNA sequencing was carried out by the dideoxy chain termination method of Sanger et al. (22). A computer-assisted sequence analysis was carried out with Seqaid (19), a software package kindly provided by Donald J. Roufa, Kansas State University.Deletion mutagenesis. Plasmids pLI210 and pLI211 containing the 2.9-kilobase (kb) insert with the lipase gene from S. aureus PS54C (11) were linearized by endonuclease digestion at the unique BamHI site and further digested with BAL 31 exonuclease to obtain various-length deletions from either end of the 2.9-kb geh insert. The digests were then phenol extracted, ethanol precipitated, ligated with T4 DNA ligase, and transformed into competent E. coli LE392 (11). Transformants were selected on L-broth plates containing 10 ,ug of chloramphenicol per ml. A panel of plasmids with various size deletions in the geh fragment was obtained from the transformants. Size estimates were made by agarose gel electrophoresis of minilysates of clones after linearization of the plasmids by restriction enzyme digestion. Alternatively, plasmids pLI210 or pLI211 were digested with restriction enzymes to delete specific sections of DNA and then religated. RESULTSDeletion mutagenesis. We reported earlier (11) that plasmids pLI210 and pLI211 carry a 2,968-base-pair (bp) DNA fragment containing the lipase gene (geh) of S. aureus PS54C which expressed lipase activity both in E. coli and S. aureus. To further localize the geh gene, various plasmids containing deletions at either end of the 2.9-kb fragment were generated. These deletions are schematically shown in Fig. 1 along with an indication of the effect of the deletion of lipase activity. Up to 500 bp could be removed from the left end of the fragment and up to 80 bp could be removed from the right end without influencing activity. Larger deletions at either end of the insert resulted in loss of enzymatic activity.ONA sequence of the geh gene. The strategy used for nucleotide sequencing is shown in Fig. 2. Each restriction fragment was subcloned into bacteriophages M13 mpl8 or ...

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Section: Resultsmentioning

confidence: 99%

Lysogenic conversion of staphylococcal lipase is caused by insertion of the bacteriophage L54a genome into the lipase structural gene

Lee¹,

Iandolo²

1986

J Bacteriol

166

103

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“…Usually the lipase produced by yeasts stays inside the cell (bound to the cell wall) and it is only secreted to the culture medium when the carbon source has been completely consumed, i.e, in the stationary phase of growth (PereiraMeirelles et al, 2000). The amount of oxygen available to the microorganisms seems to be also an important parameter, since many authors have shown the dependence of lipase productivity on system aeration and agitation (Chen et al, 1999;Elibol and Ozer, 2000;Vadehra and Harmon, 1969). The aim of this work is to evaluate the effects of perfluorodecalin addition on lipase production by Y. lipolytica, considering the improvement in oxygen supply.…”

Section: Introductionmentioning

confidence: 99%

Beneficial effects of enhanced aeration using perfluorodecalin in Yarrowia lipolytica cultures for lipase production

Amaral

Almeida

Peixoto

et al. 2006

World J Microbiol Biotechnol

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The inadequate supply of oxygen to biomass is a critical factor to the productivity of most aerobic submerged fermentations. This happens because oxygen is sparingly soluble in the aqueous media. The use of a second liquid phase of perfluorocarbon (PFC), an oxygen-carrying compound, in the culture medium can increase the availability of oxygen to the microorganisms. The effect of perfluorodecalin on Yarrowia lipolytica cultures was investigated in shake-flask cultures. It was found that the specific growth rate of Y. lipolytica, a strictly aerobic yeast, increases with increasing PFC concentration. Extracellular lipase production was increased with 20% (v/v) of PFC and agitation of 250 rev/min. It was shown that the PFC presence benefitted lipase production and not just its secretion to the extracellular medium.

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“…33 Lee e Rhee 34 conseguiram aumentar a produção de lipase a partir da bactéria Pseu domonas putida sob condições distintas de aeração e agitação. Elibol e Ozer 3 obtiveram aumento da produtividade da lipase produzida pelo fungo Rhizopus arrhizus aumentando a taxa de aeração.…”

Section: Resultsunclassified

Co-produção de lipase e biossurfactante em estado sólido para utilização em biorremediação de óleos vegetais e hidrocarbonetos

2008

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Factors Affecting Production of Staphylococcal Lipase

Cited by 35 publications

References 5 publications

Lysogenic conversion of staphylococcal lipase is caused by insertion of the bacteriophage L54a genome into the lipase structural gene

Lysogenic conversion of staphylococcal lipase is caused by insertion of the bacteriophage L54a genome into the lipase structural gene

Beneficial effects of enhanced aeration using perfluorodecalin in Yarrowia lipolytica cultures for lipase production

Co-produção de lipase e biossurfactante em estado sólido para utilização em biorremediação de óleos vegetais e hidrocarbonetos

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