IDENTIFICATION OF GABA-PRODUCING LACTIC ACID BACTERIA FROM CINCALOK FERMENTATION BASED ON THE 16S RRNA AND GAD GENES

ADHITYA NAUFAL PRIBADHI; ENDANG KUSDIYANTINI; REJEKI SITI FERNIAH

doi:10.22159/ajpcr.2021.v14i7.40881

Authors

ADHITYA NAUFAL PRIBADHI Faculty of Medicine, Wahid Hasyim University, Jl. RW Sugiarto No. 1, Mount Pati Semarang, Central Java, Indonesia.
ENDANG KUSDIYANTINI Departement of Biology, Faculty of Science and Matematics, Diponegoro University, Jl. Prof. Soedarto, SH, Tembalang Semarang, Central Java, Indonesia.
REJEKI SITI FERNIAH Departement of Biology, Faculty of Science and Matematics, Diponegoro University, Jl. Prof. Soedarto, SH, Tembalang Semarang, Central Java, Indonesia.

DOI:

https://doi.org/10.22159/ajpcr.2021.v14i7.40881

Keywords:

Cincalok, Glutamate Decarboxylase, lactic acid bacteria, 16S rRNA

Abstract

Objective: The research to identify LAB using 16S rRNA potential as high produce GABA and design primer can amplify that gad gene.

Methods: Isolation genomic form LAB, molecular identification based 16S rRNA, design primer use primer3plus, and use application serial cloner to ensure the primer can amplify to target gene.

Results: That have been carried out based on the analysis of the 16 S rRNA gene have the highest similarity to Weissella confusa strain JCM 1093 with a similarity of 98.38%, while the results of the analysis of the gad gene with several primers that have been designed are not able to amplify the gad gene owned by W. confusa.

Conclusion: The results of the analysis based on the 16S rRNA gene for lactic acid bacteria were obtained by Weissella confusa. However, for the results of identification analysis based on the gad gene, the designed primers were unable to amplify the gad gene in W. confusa.

Downloads

Download data is not yet available.

References

Achmad DI, Risa N, Puji A. Characterization of lactic acid bacteria Lactobacillus sp. RED1 from cincalok formulations. J Kim Khatulistiwa 2012; ???:???.

Pangastuti A. Species definition of procaryotes based on 16S rRNA and protein coding genes sequence. J Biol Divers 2006;7:292-6. DOI: https://doi.org/10.13057/biodiv/d070319

Kasi PD, Ariandi, Eka PT. Identification of lactic acid bacteria form Sago wastewater wirh 16S rRNA genes. Maj Ilm Biol Biosf 2019;36:35-40.

Lee KW, Shim, JM, Yao Z, Kim JA, Kim HJ, Kim JH. Characterization of a glutamate decarboxylase (GAD) from Enterococcus avium M5 isolated from Jeotgal, a Korean fermented seafood. J Microbiol Biotechnol 2017;27:1216-22. DOI: https://doi.org/10.4014/jmb.1701.01058

Siragusa S, Angelis MD, Cagno RD, Rizzello CG, Coda R, Gobbetti M. Synthesis of γ-aminobutyric acid by lactic acid bacteria isolated from a variety of Italian cheeses Appl Environ Microbiol 2007;73:7283-90. DOI: https://doi.org/10.1128/AEM.01064-07

Pribadhi AN, Kusdiyantini E, Ferniah RS. Isolation and characterization of lactic acid bacteria form fermented food cincalok as producer of γ-aminobutyric acid. Bioteknol Bios Indones 2020;7:54-62.

Walsh PS, Metzger DA, Higuchi R. Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 2013;54:506-13. DOI: https://doi.org/10.2144/000114018

Pelt-Verkuil EV, Belkum AV, Hays. Principles and technical aspect of PCR Amplification. Roterdam: Spinger; 2008. DOI: https://doi.org/10.1007/978-1-4020-6241-4

Sambrook J, Russell D. Molecular Cloning: A Laboratory Manual. 3rd ed. New York: Cold Spring Harbor Laboratory Press; 2001.

Patel J. 16S rRNA gene sequencing for bacterial pathogen identification in the clinical laboratory. Mol Diagn 2001;6:313-21. DOI: https://doi.org/10.1054/modi.2001.29158

Stackebrandt, E, Goebel BM. Taxonomic note: A place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol 1994;44:846-9. DOI: https://doi.org/10.1099/00207713-44-4-846

Hidayat P, Pancoro A. Study of molecular phulogenetics and its role in providing basic information to improve the quality of topic Orchid genetic sources. Agro Biogen 2008;4:35-40.

Lestari DA, Azrianingsih R, Hendrian H. Phylogenetic species of Annonaceae from east java collection of purwodadi botanical garden based on coding and non-coding DNA sequences. J Trop Biodivers Biotechnol 2018;3:1-7. DOI: https://doi.org/10.22146/jtbb.28308

Fusco V, Quero GM, Cho GS, Kabisch J, Meske D, Neve H, et al. The genus Weissella: Taxonomy, ecology and biotechnological potential. Front Microbiol 2015;6:1-22. DOI: https://doi.org/10.3389/fmicb.2015.00155

Thapa N, Pal J, Tamang JP. Phenotypic identification and technological properties of lactic acid bacteria isolated from traditionally processed fish products of the Eastern Himalayas. Int J Food Microbiol 2006;107:33-8. DOI: https://doi.org/10.1016/j.ijfoodmicro.2005.08.009

Miyashita M, Yukphan P, Chaipitakchonlatarn W, Malimas T, Sugimoto M, Yoshino M, et al. 16S rRNA gene sequence analysis of lactic acid bacteria isolated from fermented foods in Thailand. Microb Cult Collect 2012;28:1-9.

Komatsuzaki N, Shima J, Kawamoto S, Momose H, Kimura T. Production of γ-aminobutyric acid (GABA) by Lactobacillus paracasei isolated from traditional fermented foods. Food Microbiol 2005;22: 497-504. DOI: https://doi.org/10.1016/j.fm.2005.01.002

Borah P. Primer designing for PCR. Sci Vis 2011;11:134-6.

Sulistyaningsih E. Polymerase chain reaction (PCR): A new era of infectious disease diagnosis and management. Biomedis 2007;1:17-25.