Testing of a New Collagenase Blend for Pancreatic Islet Isolation Produced by Clostridium histolyticum

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00023001%3A_____%2F18%3A00079756" target="_blank" >RIV/00023001:_____/18:00079756 - isvavai.cz</a>

Result on the web

<a href="https://www.scirp.org/pdf/ABB_2018012615372561.pdf" target="_blank" >https://www.scirp.org/pdf/ABB_2018012615372561.pdf</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.4236/abb.2018.91003" target="_blank" >10.4236/abb.2018.91003</a>

Result language

angličtina

Original language name

Testing of a New Collagenase Blend for Pancreatic Islet Isolation Produced by Clostridium histolyticum

Original language description

Clostridium histolyticum is used for production of several proteolytic enzymes such as elastase, neutral proteases, clostripain and in particular collagenase. Besides industrial applications, collagenase has been indispensable for medical purposes including isolation of pancreatic islets for diabetes treatment. The aim of this study was to optimize the method for production and partial purification of a new collagenase blend and to test its suitability for successful pancreatic islets isolation in a rat model. Bacterial strain of C. histolyticum was sequenced for presence of the collagenase genes. Different fermentation conditions were tested and the process of collagenase extraction was modified and optimized. Samples of collagenases were taken for western blot detection, activity assessment, and ability for dissociation of pancreatic tissue. Findings indicate that concentrated trypton growth medium with pepton was the most suitable for Clostridium growth and collagenase production. Whole genome sequencing revealed two genes for collagenase and also gene for clostripain. Western blot specific detection helped to select useful production modifications. Following these modifications was also improved the yield, morphology and in vitro function of intact pancreatic islets which were finally comparable or better than those achieved using standard blends of collagenase. The results support the use of the new collagenase blend for islet isolation giving thus the opportunity to choose an alternative product. Our next steps would lead to further enzyme purification through scaling up of the production method for a wider use.

Czech name

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Czech description

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Type

J_ost - Miscellaneous article in a specialist periodical

CEP classification

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OECD FORD branch

10609 - Biochemical research methods

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2018

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Advances in bioscience and biotechnology

ISSN

2156-8456

e-ISSN

2156-8502

Volume of the periodical

9

Issue of the periodical within the volume

January 29

Country of publishing house

US - UNITED STATES

Number of pages

10

Pages from-to

26-35

UT code for WoS article

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EID of the result in the Scopus database

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