Yeast <i>Kluyveromyces lactis</i> as host for expression of the bacterial lipase: cloning and adaptation of the new lipase gene from <i>Serratia</i> sp.

Šiekštelė, Rimantas; Veteikytė, Aušra; Tvaska, Bronius; Matijošytė, Inga

doi:10.1007/s10295-015-1655-0

Cited by 11 publications

(7 citation statements)

References 31 publications

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“…pH 8,0 y a una temperatura de 37°C, también, se detectó una considerable hidrólisis del sustrato a 50°C donde se mantuvo por una hora el 60 % de su actividad relativa (Figura 4). Existen reportes de la estabilidad térmica en la lipasa extracelular de S. marcescens a 50°C [19], las lipasas mG1 y sG1 de S. marcescens expresadas en la levadura Kluyveromyces lactis mostraron actividades relativas de 60 y 29% respectivamente, a 45°C durante 30 minutos [20], mientras que la lipasa CCEI-1l, del presente estudio, tuvo una mayor actividad relativa considerando la temperatura y tiempo de incubación (50°C por 1 hora). la lipasa ECU1010 retuvo el 71% de estabilidad a 50°C durante una hora [18].…”

Section: Evaluación De La Estabilidad Térmica Y Efectos Del Ph En La unclassified

“…la mayor estabilidad de la enzima CCEI-1l a pH de 8,0 a 37 y 50°C concuerdan con las evaluaciones del efecto del pH en la producción de la lipasa por la cepa CCEI-1, donde se observó una mayor actividad de la enzima a un pH de 8,0. las lipasas mG1 y sG1 alcanzaron su mayor estabilidad en buffers con pH de 7,5 y 7,0 respectivamente, a 25°C durante 10 minutos de incubación [20], mientras que la lipasa CCEI-1l, la obtuvo a 37°C a pH 8,0 durante 1 hora de incubación.…”

Section: Evaluación De La Estabilidad Térmica Y Efectos Del Ph En La unclassified

“…En el precipitado enzimático de S. marcescens CCEI-1 se detectaron varias bandas de posibles proteínas alrededor de los 66 kDa, confirmando la presencia de una lipasa microbiana pues existen varios reportes que indican que esta enzima sintetizada por S. marcescens tiene pesos moleculares en este rango, 66,2 kDa [18,20], 65k Da [17,19] y 52 kDa [17].…”

Section: Purificación Parcial De La Enzima Y Electroforesis En Gelunclassified

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INFLUENCIA DE MICROORGANISMOS EN LA DISPONIBILIDAD DE FÓSFORO EN PLÁNTULAS DE CAFÉ (Coffea arabica)

Cisneros¹,

Franco²,

Fernandez³

et al. 2017

BSAA

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Section: Evaluación De La Estabilidad Térmica Y Efectos Del Ph En La unclassified

Section: Purificación Parcial De La Enzima Y Electroforesis En Gelunclassified

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INFLUENCIA DE MICROORGANISMOS EN LA DISPONIBILIDAD DE FÓSFORO EN PLÁNTULAS DE CAFÉ (Coffea arabica)

Cisneros¹,

Franco²,

Fernandez³

et al. 2017

BSAA

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“…E13 (Fig. 4 b) (Šiekštele et al 2015 ). Analysis of the ZS6 supernatant proteins revealed a probable lipase with a molecular weight between 63 and 75 kD in SDS-PAGE (Fig.…”

Section: Resultsmentioning

confidence: 93%

A novel Serratia sp. ZS6 isolate derived from petroleum sludge secretes biosurfactant and lipase in medium with olive oil as sole carbon source

Cheng

Liu

2018

AMB Expr

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Biosurfactants emulsify lipids, facilitating their exposure to microorganisms in water. Secretion of biosurfactant together with lipase can enhance oil assimilation by microorganisms. In this study, we show the analysis of the biosurfactant and lipase producing microbe originating from oil-sludge. The enrichment of the biosurfactant-producing ZS6 strain from the mixed culture of sludge-derived microbes in medium with olive oil as sole carbon source was monitored using the modified T-RFLP (or mT-RFLP) methodology. Phylogenetic tree analysis based on 16S rDNA sequences suggested that ZS6 belonged to a member of Serratia sp. Serratia sp. ZS6 secreted both a serrawettin-type biosurfactant and lipase in medium with olive oil as sole carbon source. By using an in-gel lipase assay followed by LC–MS/MS analysis, we identified the amino acid sequences of the ZS6 lipase, which belonged to the lipase subfamily III of the family I. Its lipolytic activity was found to be enhanced by salinity, calcium, and methanol. Together, we show that the novel isolate Serratia sp. ZS6 secretes both biosurfactant and lipase which makes it useful for applications such as in food industry wastewater treatment and biodiesel production.Electronic supplementary materialThe online version of this article (10.1186/s13568-018-0698-9) contains supplementary material, which is available to authorized users.

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“…Yeasts are highly suitable hosts for expression of heterologous proteins (Šiekštelė et al, 2015). Pichia pastoris is a well-established and widely used protein expression system for the yield of high titer of recombinant proteins and is able to secrete proteins into media (Looser et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Insight to Improve α-L-Arabinofuranosidase Productivity in Pichia pastoris and Its Application on Corn Stover Degradation

et al. 2018

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α-L-arabinofuranosidase (ARA) with enhanced specific activity and in large amounts, is needed for a variety of industrial applications. To improve ARA production with engineered methylotrophic yeast Pichia pastoris, a genetically modified ara gene from Aspergillus niger ND-1 was investigated. Through codon optimization and rational replacement of α-factor signal peptide with the native propeptide (MFSRRNLVALGLAATVSA), ARA production was improved from 2.61 ± 0.13 U/mL to 14.37 ± 0.22 U/mL in shaking flask culture (a 5.5-fold increase). Results of N-terminal sequencing showed that secreted active ARA of recombinant strain p-oARA had theoretical initial five amino acids (GPCDI) comparable to the mature sequences of α-oARA (EAEAG) and αp-oARA (NLVAL). The kinetic values have been determined for ARA of recombinant strain p-oARA (Vmax = 747.55 μmol/min/mg, Km = 5.36 mmol/L), optimal activity temperature 60°C and optimal pH 4.0. Scaling up of ARA production by p-oARA in a 7.5-L fermentor resulted in remarkably high extracellular ARA specific activity (479.50 ± 12.83 U/mg) at 168 h, and maximal production rate 164.47 ± 4.40 U/mL. In studies of corn stover degradation activity, degree of synergism for ARA and xylanase was 32.4% and enzymatic hydrolysis yield for ARA + xylanase addition was 15.9% higher than that of commercial cellulase, indicating significant potential of ARA for catalytic conversion of corn stover to fermentable sugars for biofuel production.

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Yeast Kluyveromyces lactis as host for expression of the bacterial lipase: cloning and adaptation of the new lipase gene from Serratia sp.

Cited by 11 publications

References 31 publications

INFLUENCIA DE MICROORGANISMOS EN LA DISPONIBILIDAD DE FÓSFORO EN PLÁNTULAS DE CAFÉ (Coffea arabica)

INFLUENCIA DE MICROORGANISMOS EN LA DISPONIBILIDAD DE FÓSFORO EN PLÁNTULAS DE CAFÉ (Coffea arabica)

A novel Serratia sp. ZS6 isolate derived from petroleum sludge secretes biosurfactant and lipase in medium with olive oil as sole carbon source

Insight to Improve α-L-Arabinofuranosidase Productivity in Pichia pastoris and Its Application on Corn Stover Degradation

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