Identification of GABA production from lactic acid bacteria by Thin Layer Chromatography

Abstract

Handayani R, Sulistiani, Setianingrum N. 2016. Identification of GABA production from lactic acid bacteria by Thin Layer Chromatography. Pros Sem Nas Masy Biodiv Indon 2: 208-213. The purpose of this study was to determine how to identify compounds γ-aminobutyric acid (GABA), which is produced by lactic acid bacteria. Lactic acid bacteria contribute greatly to provide functional benefits to the human body as probiotic bacteria. Probiotics are defined as live microorganisms in foodstuffs recorded in sufficient quantities and benefit gastrointestinal health. Lactic acid bacteria are also known to produce antioxidant compounds. Functions as an antioxidant shown by strains of Lactobacillus plantarum DM5. Isolates used are lactic acid bacteria isolates 478 and 515. GABA is a non-protein amino acid that is produced via decarboxylation of glutamate by the enzyme glutamate decarboxylase and distributed widely in nature among microorganisms, plants, and animals. GABA production can be determined qualitatively by Thin Layer Chromatography (TLC) or Thin Layer Chromatography (TLC) using aluminum TLC plates (Silica gel F254, Merck, Mumbai India). BAL inoculum in liquid MRS centrifuged at 1,500 x gravity for 15 minutes, then as much as 10 mL of the supernatant was spotted or dropped on TLC plates. TLC performed using a developer solution (eluent) which consists of a mixture n-butanol: acetic acid: distilled water with a ratio of 4: 1: 1 with ninhydrin dye solution with a concentration of 0.2% (w/v ethanol). When finished, the TLC plates sprayed with a solution of ninhydrin 0.5% (w/v) and then heated to 110oC for 10 minutes. GABA compounds produced by LAB isolates can be seen from the Retention factor (Rf) equal to the standard GABA used is BAL Rf = 0.47, while the standard GABA 0.48.