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090			_aB-19993
245	1	0	_aDirected evolution of Thermomyces lanuginosus lipase to enhance methanol tolerance for efficient production of biodiesel from waste grease
490	0		_vBioresource Technology, 245, p.1491-1497, 2017
520	3		_aEngineering a methanol tolerant lipase is of great importance in biodiesel production. Here, the first semi-rational method for directed enzyme evolution to enhance methanol tolerance by targeting high B-factor residues for iterative saturation mutagenesis (ISM)is reported. The best double mutant, TLL-S105C/D27R, retained 71 percent of its original activity after incubation in methanol, showing 30 percent greater methanol tolerance than TLL. TLL-S105C/D27R also displayed 27 percent higher activity over TLL. Structure modelling suggested that the increased stability of TLL-S105C/D27R was caused by the formation of a new hydrogen bond which stabilized the protein structure. E. coli (TLL-S105C/D27R)-catalyzed biotransformation of waste grease produced biodiesel in 81 percent yield in 8h, showing improvement over the 67 percent yield for E. coli (TLL), while retaining 92 percent productivity after 4 cycles of biotransformation of waste grease. The engineered TLL mutant shows high potential for commercial biodiesel production
650	1	4	_aB-FACTOR
650	1	4	_aBIODIESEL
650	1	4	_aDIRECTED EVOLUTION
650	1	4	_aMETHANOL TOLERANCE
650	1	4	_aTHERMOMYCES LANUGINOSUS LIPASE
700	1	2	_aTian, K.
700	1	2	_aTai, K.
700	1	2	_aChua, B. J. W.
700	1	2	_aLi, Z.
856	4	0	_uhttps://drive.google.com/file/d/1jRZGswgxUZNQE8rlVySk5R5BFVuFyWPW/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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