Research Journal of Microbiology1816-4935xxxx-xxxxScience International10.3923/jm.2014.95.103Bacillus
subtilis C4]]>KusumaM.P. ReddyD. Sri Rami 2201492The present investigation is envisaged to study the production of alkaline
and thermostable polygalacturonase using isolated soil Bacillus subtilis
(C4) by submerged fermentation. The extracted enzyme was subjected to partial
purification which included ammonium sulphate precipitation, dialysis, ion exchange
chromatography and affinity chromatography. The cell free supernatant polygalacturonase
activity was found to be in the 30-40% salt saturation fraction. Specific activity
of polygalacturonase increased from 33.68 to 93.05 U mL-1. Further
the fraction was loaded onto anion exchanger, DEAE Sephacel equilibrated with
20mM Tris HCl (pH 7.5 buffer). The proteins were eluted with NaCl gradient.
9.84 fold purification of the enzyme was achieved and its specific activity
was found to be 915.6 U mL-1. When subjected to affinity chromatography
the purification fold increased up to 15 fold with specific activity 13789.11
U mL-1. The homogeneity of the polygalacturonase was judged by SDS-PAGE.
It was found that PGs had a molecular weight ranging between 43 and 66 kDa.
The partially purified enzyme was characterized; the enzyme was stable at 60°C
up to 60 min at pH 9.0. The enzyme was showing 100% substrate specificity to
polygalacturonic acid. A slight activation effect was observed in the presence
of Ca2+ and Mg2+.]]>Rombouts, F.M. and W. Pilnik, 19801980pp: 227-282pp: 227-282Coutinho, P.M. and B. Henrissat,19991999pp: 3-12pp: 3-12Kashyap, D.R., S. Chandra, A. Kaul and R. Tewari,2000Bacillus sp. DT7.]]>16277282Dinu, D., M.T. Nechifor, G. Stoian, M. Costache and A. Dinischiotu,2007Aspergillus niger MIUG 16.]]>131128137Jacob, N., C. Asha Poorna and P. Prema,2008Streptomyces lydicus.]]>9966976701Semenova, M.V., S.G. Grishutin, A.V. Gusakov, O.N. Okunev and A.P. Sinitsyn,2003Aspergillus japonicus.]]>68559569Kashayp, D.R., P.K. Vohra, S.K. Soni and R. Tewari,2001Bacillus sp. DT7.]]>2312971301Miller, G.L.,195931426428Green, A. and W.L. Hughes,19951995pp: 67-90pp: 67-90Nadaroglu, H., E. Taskin, A. Adiguzel, M. Gulluce and N. Demir,2010Bacillus pumilus (P9), purification and characterisation and fruit juice application.]]>1551675175Mondal, K., P. Mehta and M.N. Gupta,2004Aspergillus niger pectinase by microwave-treated alginate.]]>33104109Stagemann, H.,19791979pp: 313-336pp: 313-336Buga, M.L., S. Ibrahim and A.J. Nok,2010Aspergillus niger (SA6).]]>989448954Roosdiana, A., S. Prasetyawan, C. Mahdi and S. Sutrisno,2013Bacillus firmus pectinase.]]>23541Kobayashi, T., Y. Hatada, N. Higaki, D.D. Lusterio and T. Ozawa et al.,1999Bacillus isolate.]]>1427145154Nagel, C.W. and R.H. Vaughn,1961Bacillus polymyxa.]]>93344352Fogarty, M.V. and C.T. Kelly,19831983pp: 131-182pp: 131-182Nakamura, N., Y. Morikawa and M. Tanaka,1971Escherichia coli Part II. substrate specificity studies.]]>35743751Martins, E.S., D. Silva, R. Da Silva and E. Gomes,2002Thermoascus aurantiacus.]]>37949954Beg, Q.K. and R. Gupta,2003Bacillus mojavensis.]]>32294304Li, Z.M., B. Jin, H.X. Zhang, Z.H. Bai, W.T. Xue and H.Y. Li,2008Bacillus gibsonii.]]>8768774