@article { ISI:000255803800012, title = {Production and single step purification of cyclodextrin glycosyltransferase from alkalophilic bacillus firmus by ion exchange chromatography}, journal = {Biochemical Engineering Journal}, volume = {39}, number = {3}, year = {2008}, month = {MAY}, pages = {510-515}, publisher = {ELSEVIER SCIENCE BV}, address = {PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}, abstract = {

Production and purification of starch digesting cyclodextrin glycosyl transferase (CGTase) from alkalophilic Bacillus firmus was investigated. Fermentation was carried out in 141 bioreactor at 28 degrees C using a medium containing dextrin, yeast extract, peptone, (NH4)H2PO4 and MgSO4.7H(2)O. The extracellular enzyme was concentrated by tangential flow ultrafiltration. The concentrated enzyme was chromatographed using DEAE-sepharose and phenyl sepharose. DEAE-sepharose could be used to purify CGTase in a single step with 23.1 fold purification and 80.6\% recovery. The enzyme obtained had homogeneity and the molecular weight was 76 kDa confirmed by SDS-PAGE. (c) 2007 Elsevier B.V. All rights reserved.

}, keywords = {alkalophilic, Bacillus firmus, CGTase, cyclodextrin, Purification}, issn = {1369-703X}, doi = {10.1016/j.bej.2007.09.020}, author = {Savergave, Laman S. and Mule, Santosh S. and Jogdand, Vitthal V. and Nene, Sanjay N. and Gadre, Ramchandra V.} } @article { ISI:000309096400004, title = {Poly(allyl glycidyl ether-co-ethylene glycol dimethacrylate) copolymer beads as support for covalent immobilization of L-aminoacylase}, journal = {Reactive \& Functional Polymers}, volume = {72}, number = {10}, year = {2012}, month = {OCT}, pages = {687-694}, publisher = {ELSEVIER SCIENCE BV}, address = {PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}, abstract = {

Porous epoxy-activated copolymer beads were synthesized as support for the covalent immobilization of Aspergillus melleus L-aminoacylase. Here, a series of copolymer bead were synthesized using either glycidyl methacrylate (GMA) or ally] glycidyl ether (AGE) as monomer units and ethylene glycol dimethacrylate (EGDM) as cross-linking agent. The effect of monomer used and the effect of amount of cross-linking agent on covalent immobilization of aminoacylase were studied. Furthermore, the effect of porogen on immobilization of aminoacylase was also evaluated. AGE-co-EGDM copolymer beads gave higher binding of aminoacylase than GMA-co-EGDM copolymer beads. AGE-co-EGDM copolymer beads synthesized with lauryl alcohol as porogen and having 150\% cross-linked density (i.e. AGE-(L)-150) gave maximum enzyme binding. Under optimum conditions, AGE-(L)-150 copolymer beads gave about 130 U/g of aminoacylase activity which corresponds to 72.24\% of activity yield. Immobilized aminoacylase showed a broader pH, higher temperature and extended storage stability. (C) 2012 Elsevier Ltd. All rights reserved.

}, keywords = {Enzyme immobilization, Epoxy-activated support, L-Aminoacylase}, issn = {1381-5148}, doi = {10.1016/j.reactfunctpolym.2012.06.012}, author = {Vaidya, Bhalchandra K. and Ingavle, Ganesh C. and Ponrathnam, S. and Nene, Sanjay N.} } @article { ISI:000300036000006, title = {Preparation of cross-linked enzyme aggregates of L-aminoacylase via co-aggregation with polyethyleneimine}, journal = {Journal of Molecular Catalysis B-Enzymatic}, volume = {74}, number = {3-4}, year = {2012}, month = {FEB}, pages = {184-191}, publisher = {ELSEVIER SCIENCE BV}, address = {PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS}, abstract = {

L-Aminoacylase from Aspergillus melleus was co-aggregated with polyethyleneimine and subsequently cross-linked with glutaraldehyde to obtain aminoacylase-polyethyleneimine cross-linked enzyme aggregates (termed as AP-CLEA). Under the optimum conditions, AP-CLEA expressed 74.9\% activity recovery and 81.2\% aggregation yield. The said method of co-aggregation and cross-linking significantly improved the catalytic stability of L-aminoacylase with respect to temperature and storage. AP-CLEA were employed for enantioselective synthesis of three unnatural amino acids (namely: phenylglycine, homophenylalanine and 2-naphthylalanine) via chiral resolution of their ester-. amide- and N-acetyl derivatives. The enantioselectivity of AP-CLEA was the highest for hydrolysis of amino acid amides; was moderate for hydrolysis of N-acetyl amino acids and was the least for hydrolysis of amino acid esters. Furthermore, AP-CLEA were found to retain more than 92\% of the initial activity after five consecutive batches of (RS)-homophenylalanine hydrolysis suggesting an adequate operational stability of the biocatalyst. (C) 2011 Elsevier B.V. All rights reserved.

}, keywords = {chiral resolution, Cross-linked enzyme aggregates, L-Aminoacylase, Polyethyleneimine, Unnatural amino acids}, issn = {1381-1177}, doi = {10.1016/j.molcatb.2011.10.003}, author = {Vaidya, Bhalchandra K. and Kuwar, Suyog S. and Golegaonkar, Sandeep B. and Nene, Sanjay N.} }