从“TapaiPulut”中筛选生物合成谷氨酸的乳酸菌

Mohd Syafarim Md Ishak, M. Ibrahim, H. Ishak
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摘要

谷氨酸(谷氨酸)是一种在人体中用于构建蛋白质的氨基酸。谷氨酸可通过谷氨酰胺合成酶转化为谷氨酰胺,通过谷氨酸脱羧酶转化为γ -氨基丁酸(GABA)。本研究的目的是筛选从“塔帕乌比”中提取的乳酸菌用于生物合成谷氨酸的潜力。利用传统发酵食品“tapaipulut”作为乳酸菌的来源。样品在含0.16%溴甲酚紫的dem - rogosa - sharpe MRS琼脂上培养,在37℃厌氧条件下培养48小时。共分离菌株25株,经形态学鉴定为LAB菌株8株。将8株菌株接种于8瓶发酵培养基(4% v/v)中,重复测定,在旋转摇床(180 rpm, 37℃,4 d)中发酵。18小时后,发酵培养基在10000 rpm(4℃)下离心10分钟,上清液在100℃下衍生10分钟。采用高效液相色谱法测定谷氨酸浓度,色谱柱为250 × 4.6 mm,流速为1.2 ml/min,荧光检测器(C18S)。从25株菌株中筛选出8株具有生产谷氨酸的能力。5株菌株(TUS 25 (18.409 μg/ml)、TUS 17 (18.437 μg/ml)、TUS 7 (17.228 μg/ml)、TUS 8 (14.033 μg/ml)、TUS 3 (7.810 μg/ml)在发酵72 h后显示出产谷氨酸的能力。其余3株菌株TUS 11 (4.894 μg/ml)、TUS 2 (0.0245 μg/ml)、TUS 20 (0.405 μg/ml)分别发酵48 h和72 h。谷氨酸在48 h和96 h时最高。本研究表明,从“tapaipulut”中提取的乳酸菌具有生产谷氨酸的能力。
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Screening of Lactic Acid Bacteria from “TapaiPulut” for Biosynthesis of Glutamic Acid
Glutamic acid (glutamate) is an amino acid used in human body to build proteins. Glutamate can be converted to glutamine via glutamine synthetase, and to γ — aminobutyric acid (GABA) via glutamate decarboxylase. The objective of this study is to screen potential of lactic acid bacteria from “tapaiubi” for biosynthesis of glutamic acid. The traditional fermented food, “tapaipulut” was used as a source of lactic acid bacteria (LAB). The sample was cultivated on DeMan-Rogosa-Sharpe MRS agar containing 0.16% bromocresol purple and placed under anaerobic condition at 37°C for 48 hours. A total of 25 strains were isolated with 8 strains were determined to be LAB based on their morphology. The 8 strains were inoculated into 8 bottles of fermentation medium (4% v/v) in duplicate determinations and fermented in rotary shaker (180 rpm, 37°C, 4 days). After 18 hours, the fermented media were centrifuged at 10000 rpm (4°C) for 10 minutes and the supernatants were derivatised at 100°C for 10 minutes. Glutamic acid concentrations were determined by using HPLC with fluorescence detector (C18S) on a 250 × 4.6 mm column at a flow rate of 1.2 ml/min. Cut of 25 strains, 8 strains were shown to have the capability of producing glutamic acid. Five strains, including (TUS 25 (18.409 μg/ml), TUS 17 (18.437 μg/ml), TUS 7 (17.228 μg/ml), TUS 8 (14.033 μg/ml), TUS 3 (7.810 μg/ml) showed the capability of producing glutamic acid after 72 hours fermentation. While the other three strains TUS 11 (4.894 μg/ml), TUS 2 (0.0245 μg/ml), TUS 20 (0.405 μg/ml) were become 48 hours and 72 hours fermentation. The highest glutamate was registered after 48 hours and 96 hours. This study shows that LAB from “tapaipulut” has the capability of producing glutamic acid.
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