Fermentation of Soybean Meal Using Co-Cultures of Lactic Acid Bacterium and Bacillus Subtilis Natto for Preparation of Feed Nutrient Additive with Anti-Obesity Activity( I )

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Project title

Code/計畫編號

MOST106-2622-E019-002-CC2

Translated Name/計畫中文名

以乳酸菌與納豆菌複合菌株發酵豆粕開發具抗肥胖活性之飼料營養添加劑( I )

Project Coordinator/計畫主持人

Guo-Jane Tsai

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=12104142

Year

2017

Start date/計畫起

01-02-2017

Expected Completion/計畫迄

01-01-2018

Bugetid/研究經費

1076千元

ResearchField/研究領域

食品科技(工)

"飼料添加劑乃為提高飼料效用與品質、促進動物發育、提高動物食用品質或其他用途而添加。飼料添加劑功能包括抗氧化、抗菌、增加動物適口性 (palatability)、改善腸胃功能及促進動物生長。近年來，動物亦開始重視肥胖問題。肥胖乃乃由於脂肪細胞内脂質合成與脂解速率之間的不平衡所致，其與脂肪細胞内賀爾蒙敏感性脂解酶(HSL)之水解油脂速率，以及脂蛋白脂解酶(LPL)產生三酸甘油酯的活性有關。已知多酚類物質（包括去醣基之大豆異黃酮）、蛋白水解胜肽、膳食纖維、prebiotics、及益生菌均具抗肥胖活性。豆粕由於蛋白質含量高且供應量充足，是近年來被研究最多的動物飼料替代蛋白原料。然而豆粕由於含有抗營養因子，尤其植酸的存在，以及Lys及Met 二種必需胺基酸較低，限制豆粕取代飼料蛋白的比例。由前期研究已獲得植酸降解率高的乳酸菌株Lactobacillus plantarum FPS 2520，且得到蛋白分解力強之納豆菌分離株N1。已知乳酸菌蛋白分解力遠低於納豆菌，但後者無法有效降解植酸及去除醣基化大豆異黃酮上之醣基。本研究主要利用乳酸菌FPS 2520及納豆菌 N1之複合菌株發酵豆粕，以開發胺基酸/胜肽含量高、植酸抗營養因子含量低、具抗肥胖活性之飼料營養添加劑。本計晝分二年進行，第一年主要探討複合菌株發酵豆粕條件。將先探討乳酸菌FPS2520與納豆菌N1各自發酵豆粕之發酵條件，以供做複合菌株發酵條件選擇之參考。複合菌株發酵將探討二株菌同時接菌發酵之模式;以及先接種N1菌，再接種FPS2520之模式。最後將各不同模式所得發酵產物，以3T3-L1細胞初步分析其抗肥胖活性。第二年將由第一年所得最佳複合菌株、以及FPS2520或N1單株菌之發酵產品，以及未發酵之豆粕為測試樣品，分析其一般組成、胺基酸組成、異黃酮含量、胜肽量、ACE抑制活性、以及3T3L-1細胞抗肥胖活性分析及對細胞内HSL及LPL活性影響。最後進行肥胖大鼠餵食試驗，探討其抗肥胖功效及可能機轉。" "Feed additives are defined as substances incorporated into diets to improve feed properties, promote animal’s growth, and increase the quality of food derived from food or others. The functions of feed additives include antioxidant and antimicrobial activities, beneficial effects on palatability and gut functions, and growth-promoting efficacy. The issue of obesity in animals is interested recently. The obesity is mainly due to the unbalance between lipolysis and synthesis of lipid in adipose tissue, which is dependent on the lipolysis rate of the hormone-sensitive lipase (HSL) and lipid production rate by lipoprotein lipase (LPL). The functional food ingredients with anti-obesity activity include dietary polyphenols (including aglycone-formed soy isoflavones), peptides from protein hydrolysates, dietary fibers, prebiotics and probiotics. Soybean meal (SBM) has the beneficial effects of high protein content and steady supply that SBM becomes as an alternative for feed protein, which has been studied frequently. However, there are two drawbacks of the lack of methionine and lysine in SBM protein, and the presence of anti-nutritional factors, especially the phytic acid, in SBM that limits its replacing percentage for animal protein in feed. A lactic acid bacterium (LAB) isolate, Lactobacillus plantarum FPS 2520, with high phytate degradation efficacy was proved, based on results from previous research project, and a strain of Bacillus subtilis natto N1 with strong protease activity was obtained in this laboratory. Based on the fact that the much weaker protein degradation ability of LABs, compared to that of natto strains, and the weaker activities of phytate degradation and isoflavone de-glyconation by natto strains, the stratage of combination of FPS 2520 and N1 will be used to ferment SBM for the preparation of a feed nutrient additive with anti-obesity activity, and with high amino acid/peptide content, low content of anti-nutritional factors. This is a two-year’s project. In the first year, the fermentation conditions for SBM fermentation by the mixed strains of FPS 2520 and N1 will be established. The fermenting conditions for SBM by individual strains will be evaluated first, which will provide the information for the second stage investigation of the conditions for mixed strains fermentation. The two models of simultaneous co-culture fermentation and lag culturing of these two strain will be investigated. The preliminary anti-obesity activity of the fermented product will be evaluated using 3T3-L1 adipocytes. In the second year, the analyses of the proximate compositions, amino acid composition, isoflavones and peptide contents, ACE inhibitory activity, and in vitro anti-obesity activity including effects on intracellular HSL and LPL activities in 3T3-L1 adipocytes will be provided on the samples of co-culture fermented product, single culture fermented product and unfermented SBM from the first year. Then, the in vivo anti-obesity activity and proposed mechanism for the tested sample will be investigated by oral feeding of the high fat diet- induced rats."

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Fermentation of Soybean Meal Using Co-Cultures of Lactic Acid Bacterium and Bacillus Subtilis Natto for Preparation of Feed Nutrient Additive with Anti-Obesity Activity( I )

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