Screening and molecular characterization of <italic>Serratia marcescens</italic> VITSD2: A strain producing optimum serratiopeptidase

doi:10.1007/s11515-013-1284-9

Front Biol

2013, Vol. 8

Issue (6) : 632-639 https://doi.org/10.1007/s11515-013-1284-9

RESEARCH ARTICLE

Screening and molecular characterization of Serratia marcescens VITSD2: A strain producing optimum serratiopeptidase

C. Subathra DEVI(

), Renuka ELIZABETH JOSEPH, Harini SARAVANAN, S. Jemimah NAINE, V. Mohana SRINIVANSAN

School of Biosciences and Technology, VIT University, Vellore, Tamil Nadu, India

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Abstract

The current work was attempted to isolate and characterize the serratiopeptidase producing Serratia sp. Among the 10 bacterial isolates 7 strains were identified as Serratia sp. Out of 7 strains one showed potent proteolytic activity and selected for further studies. Based on the morphological, biochemical and molecular characterization, the potent isolate (RH03) was identified as Serratia marcescens (GenBank accession number: KC961637) and the strain was designated as Serratia marcescens VITSD2. The production of serratiopeptidase was carried out in trypticase soya broth and the enzyme was partially purified using ammonium sulfate precipitation and dialysis. The specific activity was determined by casein hydrolysis assay and was found to be 12.00, 21.33, and 25.40 units/mg for crude, precipitated and dialysed samples. The molecular weight of the protease was determined by SDS-PAGE and it was found to be 50 kDa. The antibacterial activity of the produced serratiopeptidase showed moderate activity against Pseudomonas aeruginosa MTCC No. 4676 (12 mm) and Escherichia coli MTCC No. 1588 (15 mm).

Keywords Serratiopeptidase Serratia marcescens VITSD2 antibacterial activity

Corresponding Author(s): DEVI C. Subathra,Email:csubathradevi@vit.ac.in, subaresearch@rediffmail.com

Issue Date: 01 December 2013

Cite this article:

C. Subathra DEVI,Renuka ELIZABETH JOSEPH,Harini SARAVANAN, et al. Screening and molecular characterization of Serratia marcescens VITSD2: A strain producing optimum serratiopeptidase[J]. Front Biol, 2013, 8(6): 632-639.

URL:

https://academic.hep.com.cn/fib/EN/10.1007/s11515-013-1284-9
https://academic.hep.com.cn/fib/EN/Y2013/V8/I6/632

Tab.1 Soil samples collected from different regions of India

Tab.2 Colony morphology of the bacterial isolates

Fig.8 Red color colonies on nutrient agar.

Fig.9 Isolates showing zone of clearance on skim milk agar

Tab.3 Screening of . for Serratiopeptidase enzyme

Fig.10 Caseinolytic activity of crude enzyme (RH03)

Fig.11 Pure culture of sp. (RH03) on nutrient agar.

Tab.4 Morphological and biochemical characterization of the strain RH03

Fig.12 Phylogenetic tree of the strain VITSD2

Fig.13 (A) RNA secondary structure, (B) Mountain plot representation, (C) Base pair probabilities of Dot plot

Tab.5 Determination of SP activity and yield of enzyme from isolate VITSD2

Fig.14 SDS PAGE

Fig.15 HPLC Chromatogram of the partially purified serratiopeptidase enzyme

Tab.6 Antibacterial activity of partially purified enzyme

Fig.16 Zone of inhibition of serratiopeptidase against and (A) MTCC No: 1588 (15mm); (B) MTCC No: 4676 (12mm).

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