title: Bacillus subtillis RTSBA6 6.00, a new strain isolated from gut of Helicoverpa armigera (Lepidoptera: Noctuidae) produces chymotrypsin-like proteases.
abstract: Exploring bacterial communities with proteolytic activity from the gut of the Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) insect pests was the purpose of this study. As initial efforts to achieve this goal here we report the isolation of new Bacillus subtillis RTSBA6 6.00 strain from the gut of H. armigera and demonstrated as proteases producer. Zymographic analysis revealed 12 proteolytic bands with apparent molecular weights ranging from 20 to 185 kDa. Although some activity was detected at acidic pH, the major activity was observed at slight alkaline pH (7.8). The optimum temperature was found to be 35 °C with complete loss of activity at 70 °C. All proteases were completely inactivated by PMSF (phenylmethylsulfonyl fluoride) and TPCK (N-tosyl-l-phenylalanine chloromethyl ketone), suggesting that proteases secreted by B. subtillis RTSBA6 6.00 belong to serine proteases class with chymotrypsin-like activity. The occurrence of protease producing bacterial community in the gut of the H. armigera advocates its probable assistance to insect in proteinaceous food digestion and adaptation to protease inhibitors of host plants.
authors: Department of Biochemistry, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad 431 004, M.S., India.
authors: Ashok A
authors: AA
authors: Shinde
authors: Faiyaz K
authors: FK
authors: Shaikh
authors: Manohar V
authors: MV
authors: Padul
authors: Manvendra S
authors: MS
authors: Kachole
keywords: Bacillus subtillis RTSBA6 6.00; Helicoverpa armigera; Protease inhibitor; Serine proteases
references: Nature. 1970 Aug 15;227(5259):680-5
references: 5432063
references: Cell Mol Biol Lett. 2006;11(1):132-54
references: 16847755
references: Insect Mol Biol. 2003 Feb;12(1):85-91
references: 12542639
references: Trends Biotechnol. 1999 May;17(5):210-6
references: 10322447
references: PLoS One. 2012;7(1):e30768
references: 22292034
references: Insect Biochem Mol Biol. 2000 Jan;30(1):57-68
references: 10646971
references: J Biol Chem. 1951 Nov;193(1):265-75
references: 14907713
references: J Econ Entomol. 2000 Jun;93(3):892-6
references: 10902346
references: Comp Biochem Physiol B Biochem Mol Biol. 2001 Feb;128(2):365-75
references: 11207448
references: J Microbiol Methods. 2003 Jul;54(1):117-20
references: 12732429
references: Biotechnol Biofuels. 2009 Oct 15;2:25
references: 19832970
references: Biosci Biotechnol Biochem. 2000 Aug;64(8):1722-5
references: 10993162
references: PLoS One. 2010 May 26;5(5):e10831
references: 20520762
references: Insect Biochem Mol Biol. 2003 Feb;33(2):197-208
references: 12535678
references: Appl Environ Microbiol. 2004 Jan;70(1):293-300
references: 14711655
references: Insect Biochem Mol Biol. 2001 Mar 15;31(4-5):453-64
references: 11222955
references: Appl Microbiol Biotechnol. 1997 Sep;48(3):317-24
references: 9352674
references: J Insect Physiol. 2010 Sep;56(9):1315-24
references: 20416317
references: J Biol Chem. 2002 Oct 18;277(42):39999-40004
references: 12185078
references: Annu Rev Entomol. 2004;49:71-92
references: 14651457
references: Appl Environ Microbiol. 1994 Jun;60(6):1822-6
references: 16349275
references: Insect Biochem Mol Biol. 1997 Nov;27(11):929-44
references: 9501417
references: Can J Microbiol. 2006 Nov;52(11):1085-92
references: 17215900
references: J Biochem Biophys Methods. 1994 Apr;28(3):215-24
references: 8064116
references: Arch Insect Biochem Physiol. 2004 Mar;55(3):140-52
references: 14981658
references: Insect Biochem Mol Biol. 2010 Mar;40(3):274-83
references: 19897036
references: Environ Entomol. 2009 Aug;38(4):1078-85
references: 19689886
references: Eur J Biochem. 2002 Sep;269(17):4375-83
references: 12199717