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A highly efficient recombinant laccase from the yeast Yarrowia lipolytica and its application in the hydrolysis of biomass.

Kalyani D, Tiwari MK, Li J, Kim SC, Kalia VC, Kang YC, Lee JK - PLoS ONE (2015)

Bottom Line: The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris.It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s(-1) μM(-1)) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s(-1) μM(-1)) than other reported laccases.The standard redox potential of the T1 site of the enzyme was found to be 772 mV.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Engineering, Konkuk University, Seoul, Korea.

ABSTRACT
A modified thermal asymmetric interlaced polymerase chain reaction was performed to obtain the first yeast laccase gene (YlLac) from the isolated yeast Yarrowia lipolytica. The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris. YlLac is a monomeric glycoprotein with a molecular mass of ~55 kDa as determined by polyacrylamide-gel electrophoresis. It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s(-1) μM(-1)) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s(-1) μM(-1)) than other reported laccases. The standard redox potential of the T1 site of the enzyme was found to be 772 mV. The highest catalytic efficiency of the yeast recombinant laccase, YlLac, makes it a good candidate for industrial applications: it removes phenolic compounds in acid-pretreated woody biomass (Populus balsamifera) and enhanced saccharification.

No MeSH data available.


Related in: MedlinePlus

Dependence of the catalytic current on pH for YlLac-coated glassy carbon electrode with ABTS (○) and 2,6-DMP (■), as substrates.
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pone.0120156.g003: Dependence of the catalytic current on pH for YlLac-coated glassy carbon electrode with ABTS (○) and 2,6-DMP (■), as substrates.

Mentions: In order to investigate optimal conditions for laccase-mediated electron transfer, we evaluated the effect of pH on the catalytic current. Slow-scan voltammograms (Fig. 3) illustrate the importance of the solution pH on the catalytic current for one non-phenolic mediator (ABTS) and one phenolic mediator (2,6-DMP). Two different trends are clearly observed. For ABTS, the current decreased as the pH of the solution increased, i.e. exhibited a negative slope as pH increased. The optimum pH was around 3. On the other hand, currents recorded with 2,6-DMP exhibited a maximum value in the pH range of 3.5–5 due to proton exchange [32,33].


A highly efficient recombinant laccase from the yeast Yarrowia lipolytica and its application in the hydrolysis of biomass.

Kalyani D, Tiwari MK, Li J, Kim SC, Kalia VC, Kang YC, Lee JK - PLoS ONE (2015)

Dependence of the catalytic current on pH for YlLac-coated glassy carbon electrode with ABTS (○) and 2,6-DMP (■), as substrates.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4363317&req=5

pone.0120156.g003: Dependence of the catalytic current on pH for YlLac-coated glassy carbon electrode with ABTS (○) and 2,6-DMP (■), as substrates.
Mentions: In order to investigate optimal conditions for laccase-mediated electron transfer, we evaluated the effect of pH on the catalytic current. Slow-scan voltammograms (Fig. 3) illustrate the importance of the solution pH on the catalytic current for one non-phenolic mediator (ABTS) and one phenolic mediator (2,6-DMP). Two different trends are clearly observed. For ABTS, the current decreased as the pH of the solution increased, i.e. exhibited a negative slope as pH increased. The optimum pH was around 3. On the other hand, currents recorded with 2,6-DMP exhibited a maximum value in the pH range of 3.5–5 due to proton exchange [32,33].

Bottom Line: The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris.It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s(-1) μM(-1)) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s(-1) μM(-1)) than other reported laccases.The standard redox potential of the T1 site of the enzyme was found to be 772 mV.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical Engineering, Konkuk University, Seoul, Korea.

ABSTRACT
A modified thermal asymmetric interlaced polymerase chain reaction was performed to obtain the first yeast laccase gene (YlLac) from the isolated yeast Yarrowia lipolytica. The 1557-bp full-length cDNA of YlLac encoded a mature laccase protein containing 519 amino acids preceded by a signal peptide of 19 amino acids, and the YlLac gene was expressed in the yeast Pichia pastoris. YlLac is a monomeric glycoprotein with a molecular mass of ~55 kDa as determined by polyacrylamide-gel electrophoresis. It showed a higher catalytic efficiency towards 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (kcat/Km = 17.5 s(-1) μM(-1)) and 2,6-dimethoxyphenol (kcat/Km = 16.1 s(-1) μM(-1)) than other reported laccases. The standard redox potential of the T1 site of the enzyme was found to be 772 mV. The highest catalytic efficiency of the yeast recombinant laccase, YlLac, makes it a good candidate for industrial applications: it removes phenolic compounds in acid-pretreated woody biomass (Populus balsamifera) and enhanced saccharification.

No MeSH data available.


Related in: MedlinePlus