Purification and Characterization of Rennin Like Protease from Enterococcus faecium 1.15 Isolated from Indonesia Fermented Bakasam
Abstract
The protease enzyme produced by Lactic Acid Bacteria (LAB) with characteristics similar to rennin is called Rennin Like Protease (RLP). The purpose of the study was to obtain information on the biochemical characteristics of RLP produced by Enterococcus faecium 1.15 as a basis for the utilization of the enzyme. The first stage of the study was observation of the growth curves and the potential of the isolates to produce RLP, biochemical characterization. Rennin Like Protease (RLP) produced by Enterococcus faecium 1.15 which has been purified using Sephadex G-50 has a molecular weight of 42 kDa with biochemical characters, with an optimum temperature of 40 ℃ and pH of 4.5. The comparison between MCA/PA with a value of 3.74 which is close to the MCA value of Calf Rennet is 1.
Downloads
References
Ahmed SA, Wehaidy HR, Ibrahim OA, Hasan SA, El-Hofi MA.2016. Novel milk-clotting enzyme from Bacillus
stearothermophillus as a coagulant in UF-white soft cheese. Biocatalysis and Agricultural Biotechnology
http://dx.doi.org/10.1016/j.bcab.2016.06.011.
Azadi MA, Hemmati R, Homaei A, Khalaji-Pirbalouty V.2021. A psychrophilic caseinolytic aspartic protease
from the freshwater amphipod Gammarus bakhteyaricus for application in milk coagulation. LWT - Food
Science and Technology 153 (2022) 112536
Dini CM,Gomes E,Boscolo M,Roberto da Silva.2010.Production and characterization of a milk-clotting protease
in the crude enzymatic extract from the newly isolated Thermomucor indicae-seudaticae N31 (Milk-clotting
protease from the newly isolated Thermomucor indicae-seudaticae N31). Food Chemistry 120(2010) 87–93.
El-Bendary MA, Moharam MF, Ali TH.2007.Purification and characterization of milk-clotting enzyme produced
by Bacillus spharicus.Journal of Applied Science Research.(3) : 695-699.
El-Sayed M.M. Abdeen a, Osama A. Ibrahim, Adel M.M. Kholif.2021. Utility of Moringa oleifera waste as a
coagulant in goat soft cheese production. Heliyon 7 (2021) e07536.
Hang F, Liu P, Wang Q, Han J, Wu Z, Gao C, Liu Z, Zhang H, Chen W.2016.High milk-clotting activity
expressed by the newly isolated Paenibacillus spp.strain BD3526. Molecules, 21, 73;
doi:10.3390/molecules21010073.
Hang F, Wang Q, Hong Q, Liu P, Wu Z, Liu Z, Zhang H, Chen W.2016.Purification and characterization of a
novel milk-clotting metalloproteinase from Paenibacillus spp.BD3526. International Journal of Biological
Macromolecules (85): 547-554. http://dx.doi.org/10/j.ijbiomac.2016.01.028.
He X, Zhang W, Ren F, Gan B, Guo H.2012.Screening fermentation parameters of the milk-clotting enzyme
produced by newly isolated Bacillus amyloliquefaciens D4 from the Tibetan Plateau in China. Ann.
Microb.62:357-365.
Jiang TL, Chen J, Xue L.2007.Studi of milk-clotting mechanism of rennet-like enzyme from glutinous rice wine:
Proteolytic property and the cleavage site on k-casein.J.Dairy Sci.90:3126-3133.
Luo J, Xiao C, Zhang H, Ren F, Lei X, Yang Z, Yu Z.2017. Characterization and milk coagulating properties of
Cynanchum otophyllum Schneid. Proteases. J. Dairy Sci. 101:2842–2850.
https://doi.org/10.3168/jds.2017-13888
Mazorra-Manzano MA, Perea-Gutierrez TC, Lugo-Sanchez ME, Ramirez-Suarez JC, Torres-Llanez MJ,
Gonzales-Cordova AF, Vallejo-Cordoba B.2013.Comparison of milk-clotting properties of three plant extract.
Food Chemistry (141): 1902-1907.
Mazorra-Manzano MA, Perea-Gutierrez TC, Lugo-Sanchez ME, Ramirez-Suarez JC, Torres-Llanez MJ,
Gonzales-Cordova AF, Vallejo-Cordoba B.2013.Comparison of milk-clotting properties of three plant extract.
Food Chemistry (141): 1902-1907.
Putranto WS, Maggy TS, Harsi DK, Puspo EG, Apon ZM (2020a) A novel rennin like protease from
Lactobacillus plantarum 1.13 isolated from Indonesian fermented meat (Bakasam). Biocatal Agric Biotechnol.
https://doi.org/10.1016/j.bcab.2020.101818
Putranto WS, Mustopa AZ, Arizah K, Prastyowati A (2020b) The purifcation of rennin-like protease from
Lactobacillus paracasei isolated from ettawa goat milk. Annales Bogorienses 24:74–80.
https://doi.org/10.14203/ann.bogor.2020.v24.n2.74-80
Putranto WS, Mustopa AZ, Mamangkey J, Aritonang NJ. Characteristics of Curds With Milk Clotting Enzyme
from Indonesian Local Isolate of Lactic Acid Bacteria.2020. International Journal Of Science, Technology &
Management.79-86.
Shah MA, Mir SA, Paray MA.2014. Plant proteases as milk-clotting in cheese making:areview. J Dairy Sci and
Technol.94 (1) : 5-16.
International Journal of Science, Technology & Management ISSN: 2722 - 4015
Solanki P, Putatunda C, Kumar A, Bhatia R, Walia A.2021. Microbial proteases: ubiquitous enzymes
with innumerable uses. 3 Biotech (2021) 11:428. https://doi.org/10.1007/s13205-021-02928-z.
Wu FC, Chang CW, Shih LL.2013.Optimization of the production and characterization of milk-clotting enzymes
by Bacillus subtillis natto. Springer Plus 2: 33-42.
Zhang Y, Wang H, Tao L, Huang A.2015.Milk-clotting mechanism of Dregea sinensis Hemls.protease. J. Dairy
Sci. 98:1–9. http://dx.doi.org/10.3168/jds.2015-9851.
