Selection of acid-fast lactic acid bacterium EVL01 for inoculation in fruit and vegetable ferments to promote anti-inflammatory activity

• Main Author: 李宗益
• Other Authors: 魏佳俐
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/22rjww

碩士 === 國立嘉義大學 === 生化科技學系研究所 === 106 === Inoculation of probiotics is amongst the ways to further improve the shelf life, maintain nutritional quality and increase bioactivity. This study was aimed to develop the fruit and vegetable ferments with higher bioactivity for promoting product market competition. Acid-fast lactic acid bacteria were selected from the aging broths, which contain various fruit and vegetable ferments in Taiwan Enzyme Village Co., Ltd., by plating on MRS agars containing NaN3 and cycloheximide following anaerobic incubation, Gram stain test and inoculation in pineapple or papaya ferments supplemented without or with yuan zhi at 30°C incubation. After 16S rDNA sequencing, physiological and biochemical analyses, one of the best performed isolates was identified and renamed EVL01. In acid tolerance test, EVL01 showed the best growth in pH 4.0 MRS broth at 37℃ for 36 and 144 hours incubaction (OD600 0.98 and 1.32) than LAB a (La; OD600 0.62 and 1.11), which was the same species with EVL01 and purchased from FIRDI, and Lactobacillus rhamnosus GG (LGG; OD600 0.47 and 0.71). We further inoculated EVL01, La and LGG in pineapple ferments without or with yuan zhi. The EVL01-inoculated A and AZ referments showed the fermented effects with 3.4×107 and 1.5×108 CFU/ml live bacteria after 7 days fermentation, respectively. However, only 200 and 300 CFU/ml live bacteria in La-inoculated referments and 400 and 1.1×103 CFU/ml live bacteria in LGG-inoculated referments were detected. In the anti-inflammatory test on NO inhibition in LPS-induced RAW264.7 macrophages, EVL01-inoculated A referment showed an 11% increase as compared with the referment without any inoculation after 7 days fermentation. The study provides the acid-resistant lactic acid bacteria EVL01 with better acid-resistant ability than the same species. The increased live bacteria and anti-inflammatory activity after EVL01 inoculation in pH 3-4 of fruit and vegetable ferments may be applied for the company to develop health benefit beverage in future.