@article { ISI:000237610600013,
	title = {Purification, characterization and substrate specificity of thermostable alpha-galactosidase from Bacillus stearothermophilus (NCIM-5146)},
	journal = {Process Biochemistry},
	volume = {41},
	number = {6},
	year = {2006},
	month = {JUN},
	pages = {1311-1317},
	publisher = {ELSEVIER SCI LTD},
	type = {Article},
	address = {THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND},
	abstract = {<p>An extracellular thermostable alpha-galactosidase from Bacillus stearothermophilus (NCIM-5146) has been purified to homogeneity by chromatographic step, using Phenyl Sepharose CL-4B column. The specific activity of the enzyme was increased approximately 389-fold, from 1.03 U/mg protein to 400 U/mg protein. The molecular mass of the purified enzyme as determined by SDS-PAGE and gel filtration was 79.9 and 165.9 kDa, respectively, suggesting dimeric nature. The purified alpha-galactosidase is a non-glycosylated protein with a pI of 4.9. The pH and temperature optima for the purified enzyme are 6.5-7.0 and 65 degrees C, respectively. The alpha-galactosidase is stable over a broad pH range (3-9) and its half-life of inactivation (t(1/2)) at 70 degrees C is 30 min. The partial N-terminal sequence of alpha-galactosidase showed remarkable homology (80\% similarity) with earlier reported alpha-galactosidase from B. stearothermophilus NUB 3621. The secondary structure of the enzyme determined by circular dichroism (CD) spectroscopy exhibited alpha/beta class of protein and showed temperature induced conformational forms below and above the transition temperature. The purified enzyme showed biphasic Arrhenius plot with break point at 55 degrees C for pNPG and 50 degrees C for melibiose, raffinose and stachyose. The enzyme hydrolyzes alpha-1-3, alpha-1-4, and alpha-1-6 galactosidic linkages and not the beta-galactosidic linkages. Synthetic substrates pNPG and oNPG had lower K-m and higher K-cat as compare to natural substrates, melibiose, raffinose, and stachyose. (c) 2006 Elsevier Ltd. All rights reserved.</p>},
	keywords = {alpha-galactosidase, Bacillus stearothermophilus (NCIM-5146), Purification, Secondary structure, thermostable enzyme},
	issn = {1359-5113},
	doi = {10.1016/j.procbio.2006.01.003},
	author = {Gote, M. M. and Khan, Mohammad Islam and Gokhale, D. V. and Bastawade, Kulbhushan B. and Khire, Jayant Malhar}
}
@article { ISI:000261410200003,
	title = {Environment friendly crosslinked chitosan as a matrix for selective adsorption and purification of lipase of aspergillus niger},
	journal = {International Journal of Biological Macromolecules},
	volume = {43},
	number = {5},
	year = {2008},
	month = {DEC},
	pages = {422-425},
	publisher = {ELSEVIER SCIENCE BV},
	type = {Article},
	address = {PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS},
	abstract = {<p>Chitosan and its derivatives have been used as affinity matrices for purification of lipase from Aspergillus niger NCIM 1207. Trimellitic anhydride (TMA)-crosslinked deacetylated chitin adsorbed lipase selectively, yielding approximately 5-fold purification of the crude lipase with 70\% yield. Further 9-fold purification occurred on eluting through Sephacryl-100. These results suggest that chitosan derivatives can be used as inexpensive biopolymer matrices for the purification of lipases for industrial applications. (C) 2008 Elsevier B.V. All rights reserved.</p>},
	keywords = {Adsorption, Chitosan, Lipase, Matrix, Purification},
	issn = {0141-8130},
	doi = {10.1016/j.ijbiomac.2008.08.005},
	author = {Trimukhe, K. D. and Mahadik, N. D. and Gokhale, D. V. and Varma, Anjanikumar}
}
