© All Rights Reserved *Corresponding author. Email: wibowo.mangun@yahoo.co.id / wibowo.mangun@ui.ac.id International Food Research Journal 21(2): 527-532 (2014) Journal homepage: http://www.ifrj.upm.edu.my 1 Sulistiani, 2 Abinawanto, 3 Endang Sukara, 2 Andi Salamah, 4 Achmad Dinoto and 2* Wibowo Mangunwardoyo 1 Graduate Student, Department Biology, Faculty of Mathematic and Natural Sciences, Universitas Indonesia, Depok, Indonesia, 16424 2 Department Biology, Faculty of Mathematic and Natural Sciences, Universitas Indonesia, Depok, Indonesia, 16424 3 Research Center for Biotechnology, Indonesian Institute of Sciences (LIPI), Jl. Raya Jakarta Bogor Km 46 Cibinong, West Java, Indonesia, 16911 4 Research Center for Biology, Indonesian Institute of Sciences (LIPI), Jl. Raya Jakarta Bogor Km 46 Cibinong, West Java, Indonesia, 16911 Identifcation of lactic acid bacteria in sayur asin from Central Java (Indonesia) based on 16S rDNA sequence Abstract Sayur asin is commonly known in Indonesia, in particular Central Java, as a spontaneous fermentated mustard product by epiphytic lactic acid bacteria (LAB) from the plants. However, information regarding LAB diversity in Indonesia is lacking and fragmentary. Therefore, efforts on inventory of LAB from sayur asin in Indonesia has been conducted since April 2013 in order to determine its diversity. In the current study, LAB diversity in fve samples of sayur asin collected from Solo, Yogyakarta, and Semarang (Central Java) is reported. LAB were isolated from both of fermented mustards and fermenting liquors. Molecular identifcation of the isolates was conducted based on 16S rDNA sequence data. A total of 246 isolates of LAB was successfully isolated and identifed. The bacteria belong to 11 species, viz, Lactobacillus farciminis (15 isolates), L. fermentum (83 isolates), L. namurensis (18 isolates), L. plantarum (107 isolates), L. helveticus (1 isolate), L. brevis (1 isolate), L. versmoldensis (3 isolates), L. casei (12 isolates), L. rhamnosus (2 isolates), L. fabifermentans (3 isolates) and L. satsumensis (1 isolate). The current study revealed that L. plantarum and L. fermentum as common LAB used in sayur asin production from Central Java, Indonesia. Introduction Sayur asin has been known as traditional fermented mustard produced and consumed in many areas in Indonesia for years (Puspito and Graham, 1985). Manufacture of fermented mustard was made by addition of the salt to vegetables, allowing the growth of certain fermentable microorganisms, resulting in sensory changes bearing acidic and unique charateristics to the mustard (Chiou, 2004). Epiphytic lactic acid bacteria (LAB) of which initially amounted to slightly between 10-1000 cfu/g plant (0.001 to 1% of the total population of microorganisms) became dominant within the microorganism population in in the fermented mustard as a result of the addition on the anaerobic treatment and salt on the vegetables (Daeschel et al., 1987; Azcarate and Todd, 2010). According to Chao et al. (2009), fermented mustard is a good source for the exploration of LAB. Differences in the manufacture of fermented mustard, salt concentration, temperature during the fermentation process, and the level of anaerobiosis affect the selection of certain LAB species of in the mustard fermentation. Puspito and Graham (1985) reported that there was a difference in manufacturing techniques in the mustard fermentation in Indonesia. Several locations in central Java, such as Solo, Yogyakarta and Semarang, have traditionally been known as producers of sayur asin in Indonesia. Therefore, exploration of LAB in these areas will be important to reveal LAB diversity in Indonesian traditional fermented mustard. In the development of LAB taxonomy, several species of LAB heve been recognized as complex species, which is diffcult to distinguish by conventional techniques, such as physiological and biochemical tests. Therefore, increasing the numbers of LAB species which have phenotypic and physiological similarities using the conventional techniques often resulting in the misidentifcation (Vogel and Matthias, 2008; Nikolova et al., 2009). In addition, the conventional techniques are also time-consuming, not reproducible, and do not give accurate results (Yu et al., 2009). Development of molecular methods based on nucleotide sequence data from ribosomal DNA has complemented Keywords Diversity Fermented mustard Lactobacillus 16S rDNA Indonesia Article history Received: 1 November 2013 Received in revised form: 6 December 2013 Accepted: 12 December 2013