Gene Cloning, Purification, and Characterization of a Cold-Adapted Lipase Produced by Acinetobacter baumannii BD5

Park, Inhye; Kim, Sun‐Hee; Lee, Yong Seok; Lee, Sang Cheol; Zhou, Yi; Kim, Cheol-Min; Ahn, Soon‐Cheol; Choi, Yong-Lark

doi:10.4014/jmb.0802.130

Cited by 39 publications

(3 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Such advantages mean that recombinant proteins expressed as inclusion bodies are widely used in the commercial production of proteins [6,36]. Notably, significant numbers of cold-adapted lipases are also expressed via inclusion bodies [17,19,37,38]. Successful purification of LSK25 lipase via refolded inclusion bodies has also been reported for other cold-adapted lipases including Acinetobacter baumannii BD5 [37], Psychrobacter sp.…”

Section: Discussionmentioning

confidence: 99%

“…Notably, significant numbers of cold-adapted lipases are also expressed via inclusion bodies [17,19,37,38]. Successful purification of LSK25 lipase via refolded inclusion bodies has also been reported for other cold-adapted lipases including Acinetobacter baumannii BD5 [37], Psychrobacter sp. MBP [13] and Psychrobacter cryohalolentis K5 [38].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

New Recombinant Cold-Adapted and Organic Solvent Tolerant Lipase from Psychrophilic Pseudomonas sp. LSK25, Isolated from Signy Island Antarctica

Salwoom

Rahman

Salleh

et al. 2019

IJMS

View full text Add to dashboard Cite

In recent years, studies on psychrophilic lipases have become an emerging area of research in the field of enzymology. The study described here focuses on the cold-adapted organic solvent tolerant lipase strain Pseudomonas sp. LSK25 isolated from Signy Station, South Orkney Islands, maritime Antarctic. Strain LSK25 lipase was successfully cloned, sequenced, and over-expressed in an Escherichia coli system. Sequence analysis revealed that the lipase gene of Pseudomonas sp. LSK25 consists of 1432 bp, lacks an N-terminal signal peptide and encodes a mature protein consisting of 476 amino acids. The recombinant LSK25 lipase was purified by single-step purification using Ni-Sepharose affinity chromatography and had a molecular mass of approximately 65 kDa. The final recovery and purification fold were 44% and 1.3, respectively. The LSK25 lipase was optimally active at 30 °C and at pH 6. Stable lipolytic activity was reported between temperatures of 5–30 °C and at pH 6–8. A significant enhancement of lipolytic activity was observed in the presence of Ca2+ ions, the organic lipids of rice bran oil and coconut oil, a synthetic C12 ester and a wide range of water immiscible organic solvents. Overall, lipase strain LSK25 is a potentially desirable candidate for biotechnological application, due to its stability at low temperatures, across a range of pH and in organic solvents.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

New Recombinant Cold-Adapted and Organic Solvent Tolerant Lipase from Psychrophilic Pseudomonas sp. LSK25, Isolated from Signy Island Antarctica

Salwoom

Rahman

Salleh

et al. 2019

IJMS

View full text Add to dashboard Cite

show abstract

“…La secuencia aminoacídica inmadura es de 308 residuos, el tamaño de estas proteínas depende de la familia a la que pertenece, la especie y del espacio donde actúa. Así mismo, las lipasas extracelulares bacterianas por lo general presentan un péptido señal (Ihara et al 1991, Frenken et al 1992, Sangeetha et al 2014, Sullivan et al 1999, Park et al 2009 en la región N-terminal, reconocido por receptores de membrana y proteínas de secreción que permiten su salida al medio externo . En tal sentido, el reconocimiento de un péptido señal de 24 aa en la secuencia de la lipasa, se puede sustentar con los trabajos realizados por Alejandro-Paredes (2012) y Fernández-Jerí et al (2013) quienes describen que la lipasa de Marinobacter sp.…”

Section: Discussionunclassified

Determinación de secuencia y modelaje por homología de la lipasa de Marinobacter sp. LB

et al. 2018

View full text Add to dashboard Cite

Las lipasas de la familia I son reconocidas a nivel industrial por sus actividades catalíticas de esterificación, interesterificación y transesterificación. En esta investigación se caracterizó por análisis in silico a la lipasa de Marinobacter sp. LB aislado de las Salinas de Pilluana, San Martín. Con tal finalidad, se amplificó el gen lip mediante la reacción en cadena de la polimerasa (PCR) de punto final y la secuencia nucleotídica fue analizada in silico. Se elucidó la estructura terciaria empleando como molde a la lipasa 1EX9 de Pseudomonas aeruginosa PAO1 y se ejecutó el acoplamiento molecular con tres sustratos. El gen lip presentó 927 pb y la proteína madura, 284 aminoácidos. La lipasa posee un peso molecular de 29.99 kDa y un pI de 8.89. Asimismo, se identificaron residuos Ser78, Asp229 e His251, típicos de la triada catalítica de una lipasa de la familia I. Además, se evidenciaron once α-hélices periféricas y siete láminas-β internas. La región del bolsillo de unión y su afinidad por lípidos fue demostrada realizando acoplamientos moleculares con trioctanoina, tributirina y trioleina, con energías de -314.28, -248.11 y -215.44 kcal/mol, respectivamente; siendo los aminoácidos de interacción Asn167, Lys106, Trp172, Thr164, Ala179. En conclusión, la estructura tridimensional de la lipasa de Marinobacter sp. LB fue construida por modelamiento homólogo y validada en base a la calidad estereoquímica y el entorno de sus aminoácidos; mientras que, los análisis de acoplamiento con sustratos de lipasas permitieron evidenciar los aminoácidos que participan en el bolsillo de unión.

show abstract

A novel cold-adapted lipase from Acinetobacter sp. XMZ-26: gene cloning and characterisation

Zheng

Chu

Zhang

et al. 2011

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Acinetobacter sp. XMZ-26 (ACCC 05422) was isolated from soil samples obtained from glaciers in Xinjiang Province, China. The partial nucleotide sequence of a lipase gene was obtained by touchdown PCR using degenerate primers designed based on the conserved domains of cold-adapted lipases. Subsequently, a complete gene sequence encoding a 317 amino acid polypeptide was identified. Our novel lipase gene, lipA, was overexpressed in Escherichia coli. The recombinant protein (LipA) was purified by Ni-affinity chromatography, and then deeply characterised. The LipA resulted to hydrolyse pNP esters of fatty acids with acyl chain length from C2 to C16, and the preferred substrate was pNP octanoate showing a k(cat) = 560.52 ± 28.32 s(-1), K(m) = 0.075 ± 0.008 mM, and a k(cat)/K(m) = 7,377.29 ± 118.88 s(-1) mM(-1). Maximal LipA activity was observed at a temperature of 15°C and pH 10.0 using pNP decanoate as substrate. That LipA peaked at such a low temperature and remained most activity between 5°C and 35°C indicated that it was a cold-adapted enzyme. Remarkably, this lipase retained much of its activity in the presence of commercial detergents and organic solvents, including Ninol, Triton X-100, methanol, PEG-600, and DMSO. This cold-adapted lipase may find applications in the detergent industry and organic synthesis.

show abstract

Gene Cloning, Purification, and Characterization of a Cold-Adapted Lipase Produced by Acinetobacter baumannii BD5

Cited by 39 publications

References 0 publications

New Recombinant Cold-Adapted and Organic Solvent Tolerant Lipase from Psychrophilic Pseudomonas sp. LSK25, Isolated from Signy Island Antarctica

New Recombinant Cold-Adapted and Organic Solvent Tolerant Lipase from Psychrophilic Pseudomonas sp. LSK25, Isolated from Signy Island Antarctica

Determinación de secuencia y modelaje por homología de la lipasa de Marinobacter sp. LB

A novel cold-adapted lipase from Acinetobacter sp. XMZ-26: gene cloning and characterisation

Contact Info

Product

Resources

About