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Identification and characterization of a calcium dependent bacillopeptidase from Bacillus subtilis CFR5 with novel kunitz trypsin inhibitor degradation activity.

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Relation http://ir.cftri.com/13408/
http://dx.doi.org/10.1016/j.foodres.2017.10.049
 
Title Identification and characterization of a calcium dependent bacillopeptidase
from Bacillus subtilis CFR5 with novel kunitz trypsin inhibitor degradation
activity.
 
Creator Sharmila, G. R.
Prakash, M. Halami
Venkateswaran, G.
 
Subject 02 Bacteriology
05 Enzymes
 
Description The cereals and pulses are considered to be an important component in the food chain due to their proteinaceous
nature, but the presence of anti-nutritional factors (KTI) decreases their nutrient absorption rate. Kunitz trypsin
inhibitors (KTI) reduce the bioavailability of trypsin and are the primary cause for the existence of various
metabolic disorders. To overcome the inhibitory effect of KTI, a KTI degrading protein (BPC) was identified and
characterized from Bacillus subtilis CFR5. BPC possesses 60% identity with bacillopeptidase of B. subtilis 168. BPC
cleaves at DFVLD and DFFNNY sites of KTI which results in the formation of three inactive KTI fragments.
Subsequently, BPC was cloned in pHY300PLK and recombinant protein was used for the biochemical characterization,
sequence alignment and mutational studies. The optimal temperature and pH of the BPC was 40 °C
and 8.0, respectively. BPC is a calcium dependent metalloprotease and its activity was significantly increased by
41.2-fold in the presence of 2.5 mM Ca2+. BPC also showed moderate thermostability with the half-life of 4 h at
55 °C. Site directed mutagenesis studies in recombinant BPC revealed that mutation of Tyr49 with Phe, Tyr64
with Phe, and Pro141 to Arg affects the catalytic activity without affecting the conformation of BPC. Hence,
Tyr49, Tyr64 and Pro141 were identified as the unique residues responsible for KTI cleavage. Thus, this study leads
to the identification of a novel KTI degrading protease from B. subtilis CFR5 which cleaves and deactivates the
kunitz trypsin inhibitor.
 
Date 2018
 
Type Article
PeerReviewed
 
Format pdf
 
Language en
 
Identifier http://ir.cftri.com/13408/1/Food%20Research%20International%20103%20%282018%29%20263%E2%80%93272.pdf
Sharmila, G. R. and Prakash, M. Halami and Venkateswaran, G. (2018) Identification and characterization of a calcium dependent bacillopeptidase from Bacillus subtilis CFR5 with novel kunitz trypsin inhibitor degradation activity. Food Research International, 103. pp. 263-272. ISSN 0963-9969