Citation: Wahyuni, S.; Sarinah; Purnamasari, W.O.G.; Pato, U.; Susilowati, P.E.; Asnani; Khaeruni, A. Identification and Genetic Diversity of Amylase Producing Lactic Acid Bacteria from Brown Rice (Oryza nivara) Wakawondu Cultivar Based on 16S rRNA Gene. Fermentation 2022, 8, 691. https://doi.org/10.3390/ fermentation8120691 Academic Editor: Johannes Wöstemeyer Received: 30 October 2022 Accepted: 26 November 2022 Published: 29 November 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). fermentation Article Identification and Genetic Diversity of Amylase Producing Lactic Acid Bacteria from Brown Rice (Oryza nivara) Wakawondu Cultivar Based on 16S rRNA Gene Sri Wahyuni 1, *, Sarinah 1 , Wa Ode Gustiani Purnamasari 2 , Usman Pato 3, * , Prima Endang Susilowati 4 , Asnani 5 and Andi Khaeruni 6 1 Department of Food Science and Technology, Faculty of Agriculture, Halu Oleo University, Kendari 93232, Indonesia 2 Department of Chemistry, Faculty of Math and Natural Science, Halu Oleo University, Kendari 93232, Indonesia 3 Department of Food Science, Faculty of Agriculture, Riau University, Pekanbaru 28293, Indonesia 4 Department of Nutrition, Faculty of Health Science, Siliwangi University, Tasikmalaya 46115, Indonesia 5 Department of Aquatic Product Technology, Faculty of Fisheries and Marine Science, Halu Oleo University, Kendari 93232, Indonesia 6 Department of Plant Protection, Faculty of Agriculture, Halu Oleo University, Kendari 93232, Indonesia * Correspondence: sri.wahyuni@uho.ac.id (S.W.); usmanpato@yahoo.com (U.P.) Abstract: This study identified the genetic diversity of amylase-producing lactic acid bacteria from brown rice (Oryza nivara) Wakawondu cultivar based on the 16S rRNA gene. The ten lactic acid bacteria strains were isolated from the fermented Wakawondu rice washing water; two isolates, SBM3D and SBM4A, displayed strong amylase activity. The two isolates had the same characteristics according to both morphological and biochemical examination. The effect of fermentation time on SBM3D bacterial isolates revealed that bacterial growth at 12 h with OD values and enzyme activity, respectively, of 0.856 and 175 mU/mL, was nearly identical to the growth at 27 h with OD values of 0.886 and 176 mU/mL consecutively. Meanwhile, the bacterial isolate of SBM4A showed a significant increase in growth at 15 h with an OD value of 0.552 and enzyme activity of 99 U/mL. The maximum growth was seen at 18 h with an OD value of 0.657 and enzyme activity of 126 mU/mL. Cladogram of an SBM3D isolate with Pediococcus pentosaceus strains SL001 CP039378.1 and SRCM102740 CP028269.1 forming a sister group clad. Pediococcus pentosaceus strains SRCM102739 CP028266.1 and SRCM102738 CP028264.1 form a sister group in the cladogram of the SBM4A isolate. SBM3D and SBM4A, which are amylase-producing Pediococcus pentosaceus, can be used in food, chemical, health, and other industries. Keywords: Wakawondu; lactic acid bacteria; identification; Pediococcus pentosaceus 1. Introduction Wakawondu is one of the local upland brown rice cultivars originating from North Buton Regency, Southeast Sulawesi. Starch-rich rice washing water from Wakawondu can be utilized as a medium for fermentation. Lactic acid bacteria (LAB), which may use starch as an energy source, overgrow in the medium during the spontaneous fermentation process. LAB was isolated from the cloudy liquid in the middle of the fermented rice washing water. This process which acts as an intermediary for the formation of hormones auxin and gibberellins, contains abundant nutrients, including carbohydrates in the form of 85% starch, protein, cellulose, phosphorus, and vitamins [1]. According to Asngad et al. [2], waste is used to make syrup through fermentation with the addition of rosella plants as natural dyes. Presently, rice washing water has been used as a source of isolates to obtain lactic acid bacteria (LAB) and bioethanol. Therefore, this process produces more useful products [3]. Fermentation 2022, 8, 691. https://doi.org/10.3390/fermentation8120691 https://www.mdpi.com/journal/fermentation