eprintid: 9933
rev_number: 11
eprint_status: archive
userid: 1305
dir: disk0/00/00/99/33
datestamp: 2017-03-31 10:03:03
lastmod: 2017-06-22 08:41:24
status_changed: 2017-03-31 10:03:03
type: article
metadata_visibility: show
contact_email: Library-ICRISAT@CGIAR.ORG
creators_name: Akbar, S M
creators_name: Sharma, H C
icrisatcreators_name: Akbar, S M
icrisatcreators_name: Sharma, H C
affiliation: ICRISAT (Patancheru)
country: India
title: Alkaline serine proteases from Helicoverpa armigera: potential candidates for industrial applications
ispublished: pub
subjects: h1
subjects: s2.7
divisions: CRPS2
full_text_status: restricted
keywords: Alkalophilic, Chymotrypsin, Halophilic, Helicoverpa armigera, Industrial application, Serine protease, Trypsin, Thermostable
note: We thank the staff of EntomologyUnit, International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), for
providing support for insect rearing. This study was funded by Department of Science and Technology – Scientific and
Engineering Research Board (DST-SERB), New Delhi, India.
abstract: We characterized trypsin- and chymotrypsin-like serine alkaline proteases from cotton bollworm, Helicoverpa armigera, for their probable potential application as additives in various bio-formulations. Purification was achieved by using hydroxylapatite, DEAE sephadex and CM sephadex columns, which resulted in increased enzyme activity by 13.76- and 14.05-fold for trypsin and chymotrypsin, respectively. Michaelis–Menten constants (Km) for substrates of trypsin and chymotrypsin, BApNA and SAAPFpNA, were found to be 1.25 and 0.085 mM, correspondingly. Fluorescent zymogram analysis indicated the presence of five trypsin bands with molecular masses of ∼21, 25, 38, 40, and 66 kDa and two chymotrypsin bands with molecular masses of ∼29 and 34 kDa in SDS-PAGE. The optimum pH was 10.0 and optimum temperature was 50°C for proteolytic activity for the purified proteases. The proteases were inhibited by synthetic inhibitors such as PMSF, aprotonin, leupeptin, pefabloc, and antipain. TLCK and TPCK inhibited about 94 and 90% of trypsin and chymotrypsin activity, respectively, while EDTA, EGTA, E64, pepstatin, idoacetamide, and bestatin did not affect the enzymes. The purified enzymes exhibited high stability and compatibility with metal ions; oxidizing, reducing, and bleaching agents; organic solvents; and commercial detergents. Short life cycles, voracious feeding behavior, and production of multiple forms of proteases in the midgut with rapid catalytic activity and chemostability can serve H. armigera as an excellent alternative source of industrially important proteases for use as additives in stain removers, detergents, and other bio-formulations. Identification of enzymes with essential industrial properties from insect species could be a bioresource.
date: 2017-01
date_type: published
publication: Archives of Insect Biochemistry and Physiology
volume: 94
number: 1
publisher: Wiley
pagerange: 1-15
id_number: 10.1002/arch.21367
refereed: TRUE
issn: 0739-4462
official_url: http://dx.doi.org/10.1002/arch.21367
related_url_url: https://scholar.google.co.in/scholar?q=Alkaline+serine+proteases+from+Helicoverpa+armigera%3A+potential+candidates+for+industrial+applications&btnG=&hl=en&as_sdt=0%2C5
related_url_type: pub
funders: Department of Science and Technology – Scientific and Engineering Research Board (DST-SERB), New Delhi, India
citation:   Akbar, S M and Sharma, H C  (2017) Alkaline serine proteases from Helicoverpa armigera: potential candidates for industrial applications.  Archives of Insect Biochemistry and Physiology, 94 (1).  pp. 1-15.  ISSN 0739-4462     
document_url: http://oar.icrisat.org/9933/1/Akbar_et_al-2017.pdf