pISSN 2302-1616, eISSN 2580-2909 Vol 10, No. 1, June 2022, pp. 53-65 Available online http://journal.uin-alauddin.ac.id/index.php/biogenesis DOI https://doi.org/10.24252/bio.v10i1.27349 Copyright © 2022. The authors. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/ ) Identification of lactic acid bacteria and quality parameter of Tempeh obtained from red kidney beans (Phaseolus vulgaris) and cowpeas (Vigna unguiculata) Gergonius Fallo 1 , Yuni Sine 1* 1 Department of Biology, Faculty of Agriculture, Universitas Timor Sasi, Kefamenanu, North Central Timor, East Nusa Tenggara, Indonesia. 85611 *Email: sineyuni@gmail.com ABSTRACT. Tempeh fermentation generally uses soybeans as a substrate. However, the availability of soybeans in Indonesia, especially in the province of East Nusa Tenggara, is inadequate, so alternative fermentation substrates are important to consider. Red beans and cowpeas grow abundantly in East Nusa Tenggara. Therefore, researchers are interested in conducting research using local beans as a substrate for tempeh fermentation. This study aims to utilize red beans and cowpeas as substrates for tempeh fermentation with proximate analysis and isolate and identify lactic acid bacteria from bean soaking water, considering the important role of LAB in LAB the tempeh fermentation process. Red beans and cowpeas were employed, MRSA and MRSB media, test media, commercial tempeh starter, and LAB. Before the study started, traditional tempeh was prepared using a commercial starter ( Rhizopus sp.). Red peanuts and cowpeas tempeh were firstly tested using the AOAC method, secondly isolated using the spread plate method, then purified and lastly characterized and molecularly identified using PCR method and DNA sequence analysis to obtain a phylogeny tree. Based on the proximate analysis results for water content, cowpea seeds increased during soaking to fermentation, which was 6.81% to 17.41%. For the dry matter of the two samples, red beans, 7.16% to 22.22%, experienced a decrease in cowpea from 93.19% to 82.59%. Kidney beans 92.84% to 77.78%. The ash content of cowpea is 4.66% to 4.16%. red beans 4.44% to 2.41%, cowpea crude fiber 6.04% to 4.67%, red beans 5.32% to 3.75%. cowpea carbohydrates 57.049% to 40.389%, red beans 54.364% 38.474%. The protein content of cowpeas was 30.870% to 36.210% in tempeh, red beans 33.173% to 36.062% in tempeh. Cyanide levels in cowpeas were 0.043% to 0.010%, red beans were 0.054% to 0.011% in tempeh. The lactic acid bacteria that were successfully isolated and identified were bacteria with isolate code KM2 related to Enterococcus faecium and KN2 isolate Lactiplantibacillus plantarum, lactic acid bacteria function to lower the pH in the soaking water and prepare a suitable pH for the growth of Rhizopus mold. Keywords: crude fat content; molecular identification; phylogenetic tree; proximate analysis; Tempeh fermentation Article History: Received 11 February 2022; Received in revised form 14 April 2022; Accepted 13 May 2022; Available online 30 June 2022. Ver: Pre-Press How to Cite This Article: Fallo G, Sine Y. 2022. Identification of lactic acid bacteria and quality parameter of Tempeh obtained from red kidney beans (Phaseolus vulgaris) and cowpeas (Vigna unguiculata). Biogenesis: Jurnal Ilmiah Biologi. vol 10(1): 53–65. doi: https://doi.org/10.24252/bio.v10i1.27349. INTRODUCTION Tempeh is a good source of nutrition because it contains protein, carbohydrates, essential fatty acids, vitamins, and minerals and is a source of vegetable protein. Tempeh contains various nutrients needed by the body, such as protein, fat, carbohydrates, and minerals (Winarno et al., 2021). Several studies have shown that tempeh is easier to digest than soybeans. Fermented tempeh is a product that has a better shape, texture and taste and is more easily absorbed and utilized by the body (Galand et al., 2021). The moulds that grow on soybeans hydrolyze complex compounds into simple compounds that humans easily digest. Tempeh which the Indonesian people commonly known, uses soybean (Glycine max) as raw material. Tempeh contains unique biochemical properties, including particular amino acids (isoleucine, leucine, lysine, tryptophan and methionine). Tempeh is becoming more widely consumed on a global scale. Tempeh can also be made from various legumes (Radita et al., 2017). Soy tempeh is fermented in a controlled manner using a mould Rhizopus oligosporus. Apart from the mould R. oligosporus, other microorganisms are also known to participate in the tempeh fermentation process (Radita et al., 2017). Tempeh fermentation consists of two stages, including lactic acid fermentation, which occurs naturally during the soaking process by lactic acid microbiota and Rhizopus fungal fermentation (Radita et al., 2017).