Prospects for Primary Design Study of Protease Encoding Genes for Recombinant Protease Enzymes - A Bibliographic and Journal Review

DOI

10.2991/978-94-6463-457-0_2How to use a DOI?

Keywords

Specific Primer Design Software; Protease Genes; Recombinant Protease Enzymes; Cloning and Overexpression; Bacillus sp. HSFI-3

Abstract

Protease enzymes in the health sector are very important, especially as anticoagulant, antithrombotic agents, as diagnostic tools and plasmin standards. It is not surprising that the need for protease enzymes in the world is very high, including in Indonesia. The supply of protease in Indonesia still depends on imports so its availability is limited. One way to overcome this problem is to produce protease enzymes from Indonesia’s biological resources. Previous research reported that the bacteria Bacillus sp. HSFI-3 isolated from fermented digestive organs of Holothurian scabra sand sea cucumbers (West Nusa Tenggara) has proteolytic, fibrinolytic and thrombolytic characteristics. Bacillus sp. HSFI-3 is a wild type bacteria. The challenge with wild type bacteria as protease producers is that the gene promoter is not always on so it does not always express protease, the enzyme activity is low and it is not pure. Strategies need to be implemented to overcome this, one of which is cloning and overexpressing proteases for scale up efforts from wild type Bacillus sp bacteria. HSFI-3. The steps to carry out cloning and overexpression require the full-length gene sequence encoding the protease enzyme from Bacillus sp. HSFI-3. Full-length gene sequences can be obtained from the results of Whole Genome Sequencing (WGS) of Bacillus sp. HSFI-3 amplified from specific primer design. Full-length gene encoding the protease enzyme from Bacillus sp. After obtaining HSFI-3, it was then continued by amplifying the full-length gene using in silico PCR and in vitro PCR with specific primer designs based on the WGS sequence results. This journal review summarizes the types of primer design software specific to protease genes throughout the world in the last decade. This research specifically targets the software used in the design of specific primers for the full-length protease gene. The aim is to obtain the most appropriate software recommendations regarding specific primer design for the full-length protease encoding gene from Bacillus sp. HSFI-3. Based on the results of the journal review, all the software contained in this study only amplifies fragments. Benchling software (https://www.benchling.com/) is capable of designing primers for full-length genes. So we recommend this software for the design of full-length gene specific primers for the protease gene of Bacillus sp. HSFI-3 is an effort to scale up protease enzymes so that the need for protease enzymes in Indonesia is met.

Copyright

© 2024 The Author(s)

Open Access

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TY  - CONF
AU  - Rima Agnes Widya Astuti
AU  - Nanik Rahmani
AU  - Dewi Seswita Zilda
AU  - Sri Darmawati
AU  - Stalis Norma Ethica
PY  - 2024
DA  - 2024/07/22
TI  - Prospects for Primary Design Study of Protease Encoding Genes for Recombinant Protease Enzymes - A Bibliographic and Journal Review
BT  - Proceedings of the 2nd Lawang Sewu International Symposium on Health Sciences: Medical Laboratory Technology (LSISHS-MLT 2023)
PB  - Atlantis Press
SP  - 5
EP  - 15
SN  - 2468-5747
UR  - https://doi.org/10.2991/978-94-6463-457-0_2
DO  - 10.2991/978-94-6463-457-0_2
ID  - Astuti2024
ER  -