本文關(guān)鍵詞： 乳酸菌 益生特性 降血糖 篩選 發(fā)酵乳　出處：《陜西科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：高血糖可導(dǎo)致糖尿病,對(duì)人體的危害很大。乳酸菌已被公認(rèn)為對(duì)人體有益的可食用菌群,具有多種功能性,目前還在不斷研究其新的功能性,關(guān)于乳酸菌降血糖作用及其健康產(chǎn)品的開發(fā)研究已逐漸成為熱點(diǎn)。本研究以從陜西、甘肅等地所采集的傳統(tǒng)發(fā)酵食品中分離純化的SL01、SL02、SL03、SL04、SL05、SL06、SL07、SL08、SL09、SL10、SL11、SL12、SL13、SL14、SL15、SL16、SL17、SL18等18株乳酸菌為實(shí)驗(yàn)菌株,首先研究其對(duì)胃液和腸液的耐受性及產(chǎn)酸能力,其次分別對(duì)其菌懸液、發(fā)酵上清液、細(xì)胞破碎物上清液與降血糖相關(guān)的性能,即:對(duì)α-葡萄糖苷酶、α-淀粉酶抑制性、還原能力、對(duì)自由基DPPH和超氧陰離子(O2-·)的清除能力等進(jìn)行研究,篩選出綜合能力較強(qiáng)的乳酸菌,并以此為菌株,對(duì)初步制備的發(fā)酵乳降血糖作用相關(guān)指標(biāo)進(jìn)行了分析檢測(cè)。主要研究結(jié)果如下:乳酸菌的益生特性研究:通過(guò)對(duì)18株乳酸菌的形態(tài)特征,生長(zhǎng)曲線,溶鈣圈、產(chǎn)酸量及其對(duì)人工胃液和人工腸液的耐受性等方面的研究。結(jié)果表明:18株乳酸菌中16種為桿菌,2種為球菌;18株菌在24h后均可達(dá)到生長(zhǎng)旺盛期;均具有產(chǎn)酸性能,其中溶鈣圈直徑在1.35cm~2.60cm,產(chǎn)酸量為6.84g/Kg~19.98g/Kg;18株菌株對(duì)胃腸液耐受能力相差較大,胃液3h存活率20%以下有2株,20%至60%的有13株,60%以上有3株分別為菌株SL03、SL02和SL07;腸液6h存活率在20%以下有2株,存活率在20%至75%有13株,75%以上有3株分別為菌株SL03、SL09和SL18。乳酸菌與降血糖作用相關(guān)的指標(biāo)研究:分別研究了18株菌的菌懸液、發(fā)酵上清液、細(xì)胞破碎物上清液對(duì)α-葡萄糖苷酶和α-淀粉酶的抑制性、還原能力、對(duì)自由基DPPH和超氧陰離子(O2-·)的清除能力,結(jié)果表明其對(duì)與降血糖作用相關(guān)的指標(biāo)基本都具有一定的作用,具體為:菌懸液對(duì)α-葡萄糖苷酶均具有抑制性,抑制率從5.71%至37.48%,且總體高于發(fā)酵上清液和細(xì)胞破碎物上清液,對(duì)α-淀粉酶無(wú)抑制性,菌懸液有17種具有還原能力,還原濃度從9.67μmol/L至46.13μmol/L,對(duì)自由基O2-·有清除力的有4種,清除率從3.50%至23.09%,對(duì)自由基DPPH均具有清除力,清除率從9.73%至36.63%;發(fā)酵上清液對(duì)α-葡萄糖苷酶均具有抑制性,抑制率從1.55%至24.62%,15種對(duì)α-淀粉酶具有抑制性,抑制率從5.14%到33.85%,發(fā)酵上清液均具有還原能力,還原濃度從44.75μmol/L至105.25μmol/L,總體高于菌懸液和細(xì)胞破碎物上清液,對(duì)自由基O2-·和自由基DPPH均具有清除能力,O2-·清除率從2.97%至34.18%,自由基DPPH清除率從18.83%至82.92%,總體高于菌懸液和細(xì)胞破碎物上清液;細(xì)胞破碎物上清液對(duì)α-葡萄糖苷酶均具有抑制性,抑制率從0.31%至25.47%,對(duì)α-淀粉酶無(wú)抑制性,細(xì)胞破碎物上清液均具有還原能力,還原濃度從1.93μmol/L至35.64μmol/L,14種對(duì)自由基O2-·有清除力,清除率從0.74%至14.97%,對(duì)自由基DPPH均具有清除力,清除率從4.45%至43.53%。乳酸菌的鑒定:對(duì)18株菌中降血糖作用指標(biāo)較優(yōu)的SL01、SL02、SL03、SL05、SL12和SL18等6株菌進(jìn)行了進(jìn)一步鑒定,經(jīng)形態(tài)學(xué)鑒定和16S rRNA基因測(cè)序結(jié)果表明,SL01為L(zhǎng)actobacillus sp.,SL02為L(zhǎng)actobacillus casei,SL18為Enterococcus faecium,SL03、SL02和SL12均為L(zhǎng)actobacillus reuteri。乳酸菌發(fā)酵乳的制備及其與降血糖作用相關(guān)指標(biāo)的研究:以上述6株乳酸菌為發(fā)酵菌株進(jìn)行發(fā)酵乳的制備,并分析檢測(cè)發(fā)酵過(guò)程中與降血糖相關(guān)的指標(biāo)變化,結(jié)果為:6株菌制備的發(fā)酵乳均具有不同程度的降血糖活性,不同溫度對(duì)于發(fā)酵乳的降血糖作用相關(guān)指標(biāo)活性影響不同。較優(yōu)發(fā)酵條件為SL01發(fā)酵乳溫度37℃發(fā)酵72h,SL05發(fā)酵乳溫度41℃發(fā)酵72h,SL02和SL18發(fā)酵乳溫度37℃發(fā)酵56h,SL12發(fā)酵乳溫度41℃發(fā)酵48h,SL03發(fā)酵乳溫度37℃發(fā)酵64h;將6株菌制備的發(fā)酵乳等量混合后,其降血糖活性顯著增強(qiáng),其中對(duì)α-葡萄糖苷酶和α-淀粉酶的抑制率分別為25.68%和65.27%,還原能力為152.75μmol/L,O2-·清除率為40.82%,自由基DPPH的清除率為65.29%,冷藏21d之后,活菌數(shù)為5.3×107CFU/mL,酸度在109 oT以下,α-葡萄糖苷酶和α-淀粉酶的抑制率分別保持在20.58%和61.23%,還原濃度保持在143.38μmol/L,O2-·清除率保持在30.97%,DPPH清除率保持在52.62%,冷藏后混合乳降血糖作用相關(guān)指標(biāo)活性仍保持了較高值。本文通過(guò)乳酸菌的篩選、益生特性研究、降血糖作用相關(guān)指標(biāo)研究及其發(fā)酵乳的制備與降血糖作用研究,可為進(jìn)一步研究開發(fā)新型功能性乳酸菌發(fā)酵乳提供一定的理論與實(shí)驗(yàn)依據(jù)。
[Abstract]:High blood sugar can lead to diabetes, great harm to the human body. The lactic acid bacteria has been recognized as beneficial to human edible flora, with a variety of functions, is continuing to study the new features of the development of research on the hypoglycemic effect of lactic acid bacteria and health products has gradually become the hot spot in this study from Shaanxi. Separation and purification of traditional fermented food, the acquisition of Gansu and other places in SL01, SL02, SL03, SL04, SL05, SL06, SL07, SL08, SL09, SL10, SL11, SL12, SL13, SL14, SL15, SL16, SL17, SL18 and other 18 strains of lactic acid bacteria strains, we first study its tolerance to gastric juice and intestinal juice and acid production ability, followed by the bacterial suspension and supernatant, performance, broken cell supernatant associated with lowering blood sugar: the alpha glucosidase, alpha amylase inhibition, reducing power, DPPH free radical and superoxide anion (O2- -) of the scavenging capacity etc. Study on screening of lactic acid bacteria, strong comprehensive ability, and as a strain of fermented milk, preliminary preparation reduced blood glucose related indexes were detected. The main results are as follows: Study on probiotic characteristics of lactic acid bacteria: the morphological characteristics of 18 strains of lactic acid bacteria growth curve, dissolved calcium circle, study acidity and tolerance on artificial gastric juice. The results showed that the 18 strains of lactic acid bacteria in 16 kinds of bacteria, 2 for Staphylococcus aureus 18 strains; can achieve growth in 24h after Washings; had acid production, which dissolved calcium circle diameter in 1.35cm~2.60cm acid production was 6.84g/Kg~19.98g/Kg; 18 strain difference on gastrointestinal tolerance, gastric 3H survival rate below 20% and 2 strains, 20% to 60% of the 13 strains, 3 strains were more than 60% strains SL03, SL02 and SL07; intestinal 6h survival rate below 20% in 2 strains, the survival rate in 20% to 75% in 13 strains, More than 75% of 3 strains were strains SL03, SL09 and SL18. index to study the hypoglycemic effect of lactic acid bacteria and related: 18 strains of bacterial suspension and supernatant, broken cell supernatant of alpha glucosidase and alpha amylase inhibition, reduction ability of free radical DPPH (O2- -) and superoxide anion scavenging ability, the results showed that the correlation with the hypoglycemic effect of basic indicators has a certain role, in particular: the bacterial suspension showed inhibition of alpha glucosidase inhibitory rate, from 5.71% to 37.48%, and was higher than that of fermentation supernatant and cell lysate supernatant. The alpha amylase inhibiting bacterial suspension, 17 has the ability to restore, restore the concentration from 9.67 mol/L to 46.13 mol/L, the O2- radical scavenging capacity of 4, the clearance rate from 3.50% to 23.09%, with scavenging DPPH free radical scavenging rate from 9.73% to 36.. 63%; fermentation supernatant has inhibition of alpha glucosidase inhibitory rate, from 1.55% to 24.62%, with 15 inhibition of alpha amylase inhibition rate from 5.14% to 33.85%, the fermentation supernatant showed good reducing ability, reducing the concentration from 44.75 mol/L to 105.25 mol/L, the overall high in suspension and the cell supernatants were broken, with the radical scavenging activity of O2- and DPPH radical, O2- radical scavenging rate from 2.97% to 34.18%, DPPH free radical scavenging rate from 18.83% to 82.92%, higher than the overall bacterial suspension and supernatant of cell disruption; broken cell supernatant on alpha glucosidase inhibitory have. The inhibition rate from 0.31% to 25.47%, the alpha amylase inhibiting cell supernatant were broken, has the ability to restore, restore the concentration from 1.93 mol/L to 35.64 mol/L, 14 kinds of O2- radical scavenging capacity, scavenging rate from 0.74% to 14.97%, with the free radical DPPH There are clear, the clearance rate of 43.53%. to the identification of lactic acid bacteria from 4.45%: the hypoglycemic effect index of optimal SL01, 18 strains SL02, SL03, SL05, SL12 and SL18 and other 6 strains were further identified by morphological identification and 16S rRNA gene sequencing results showed that SL01 Lactobacillus sp., SL02 Lactobacillus casei, SL18 Enterococcus faecium, SL03, SL02 and SL12 are Lactobacillus reuteri. of lactic acid bacteria fermented milk preparation and hypoglycemic effect of relevant indicators: the 6 strains of lactic acid bacteria fermentation strains were fermented milk preparation, and analysis of changes in indicators of blood glucose detection in the fermentation process and the lower correlation results: 6 strains of Lactobacillus fermentation were prepared with hypoglycemic activity in different degree, relevant indicators of the hypoglycemic effect of different temperature on the activity of fermented milk have different effects. The optimal fermentation conditions for SL01 fermented milk fermentation temperature 37 C 7 2H, SL05 fermented milk fermentation temperature 41 72h, SL02 and SL18 in fermented milk temperature 37 56h fermentation, SL12 fermented milk fermentation temperature is 41 DEG 48h, SL03 fermented milk fermentation temperature is 37 DEG 64H; 6 strains preparation of fermented milk mixture, its hypoglycemic activity was significantly enhanced and the inhibition of alpha glucosidase and alpha amylase were 25.68% and 65.27%, reducing capacity of 152.75 mol/L, O2- clearance rate was 40.82%, the free radical scavenging rate of DPPH was 65.29%, refrigerated after 21d, number of live bacteria is 5.3 * 107CFU/mL, pH below 109 oT, inhibition of alpha glucosidase and alpha amylase rate was kept at 20.58% and 61.23%, the reduction of concentration is kept at 143.38 mol/L, O2- removal rate remained at 30.97%, the clearance rate of DPPH remained at 52.62%, after cold storage of mixed milk hypoglycemic activity index still has high value. Through the screening of lactic acid bacteria, study on probiotic characteristics drop Studies on related indicators of blood glucose, preparation and hypoglycemic effect of fermented milk can provide theoretical and experimental basis for further research and development of new functional lactic acid bacteria fermented milk.

【學(xué)位授予單位】：陜西科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TS201.3

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