專家同意對外發(fā)布的乳酸菌會議大會報告何國慶2014

1、防?。庖呙阜啦。庖呙赴肴樘擒彰负啺肴樘擒彰负啴a(chǎn)半乳糖苷酶乳酸菌的篩選與鑒半乳糖苷酶的提純及全細胞酶與生物量的相關(guān)生物膜的誘1.1 -半乳糖苷酶分類學(xué)地EC（Enzyme）EGlyosidase data1.1 -半乳糖苷酶分類學(xué)地EC（Enzyme）EGlyosidase dataEnzymes半乳糖苷酶來索引嗜堿芽孢桿菌（Bacillus （嗜酸乳桿菌（Lactobacillus 嗜熱古細菌（Pyrococcus 多形擬桿菌（Bacteroides 脆弱類擬桿菌（Bacteroides se: 糖苷水解酶位乳桿菌雙歧桿菌乳桿菌雙歧桿菌其他：乳球菌（Lactococcus）、片球菌(Ped

2、iococcus)消除豆粕類飼料中抗營養(yǎng)因消除豆粕類飼料中抗營養(yǎng)因食品中的應(yīng)半乳糖苷酶在食品中的應(yīng)用，主要是用于豆乳的發(fā)酵以及甜菜制糖工業(yè)法布里氏?。‵abry 半乳糖苷酶與BO血型轉(zhuǎn)顯色圈 （3748樣品準（37245PYSE半選培養(yǎng)基初（374800酒 糟 酸菜酒 曲筍干株篩（3748.株純養(yǎng)1824 h）-XA1R XB4R XC7R XD6R XF5R XH4B -顯色時間 上清液所占總酶活比率顯色圈顯色時間 顯色圈 （3748樣品準（37245PYSE半選培養(yǎng)基初（374800酒 糟 酸菜酒 曲筍干株篩（3748.株純養(yǎng)1824 h）-XA1R XB4R XC7R XD6R XF5R

3、 XH4B -顯色時間 上清液所占總酶活比率顯色圈顯色時間 唾液乳桿唾液唾液乳桿唾液Lactobacillussa varius乳球230.51m的短桿菌乳酸片球菌則是直徑約 12 m 的球菌。Lactobacillushordei Lactobacillus maliLactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus ceti LactobacillussaerimneriLactobacillus LactobacillussalivariusXA1R Lactobacillus salivarius

4、Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillusmanihotivorans Lactobacillus caseiL.salivariusLactobacilluscurvatus Lactobacillus sakeiLactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus L.salivariusLactobacillus Lactob

5、acillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus plantarum Lactobacillusmalefermentans Lactobacillus parakefiriilacticiLactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Pediococcus Pediococcus acidilactici Pediococcus Pediococcus Pediococcus Pediococc

6、us parvulus PediococcusdamnosusPediococcus Lactobacillushordei Lactobacillus maliLactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus ceti LactobacillussaerimneriLactobacillus LactobacillussalivariusXA1R Lactobacillus salivarius Lactobacillus Lactobacillus Lactobacillus Lactobacillu

7、s Lactobacillus Lactobacillusmanihotivorans Lactobacillus caseiL.salivariusLactobacilluscurvatus Lactobacillus sakeiLactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus L.salivariusLactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lact

8、obacillus plantarum Lactobacillusmalefermentans Lactobacillus parakefiriilacticiLactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Lactobacillus Pediococcus Pediococcus acidilactici Pediococcus Pediococcus Pediococcus Pediococcus parvulus PediococcusdamnosusPediococcus 比率 864-2-0凍

9、融法超聲法脲酶法-溶菌 酶法對照酶活比率 864-2-0凍融法超聲法脲酶法-溶菌 酶法對照酶活有0洗脫OD280nm項目粗超柱層蛋白質(zhì)濃（U/g 純化倍有0洗脫OD280nm項目粗超柱層蛋白質(zhì)濃（U/g 純化倍約樣品說明Ur:約樣品說明Ur:4MCr: Sf:超濾酶Sc:554433221100(A) Temperautre (B) pH Catalyst activity Pure enzyme Whole cell enzymePure enzyme Whole cell enzyme554433221100(A) Temperautre (B) pH Catalyst activity

10、Pure enzyme Whole cell enzymePure enzyme Whole cell enzyme3 檸檬酸緩沖液pH 離子強度0.15 52A:pH3 檸檬酸緩沖液pH 離子強度0.15 52A:pHB: temperature Catalyst activity3 3反應(yīng)溫度pNPG Km值線性線性9876y=0.5536x反應(yīng)溫度pNPG Km值線性線性9876y=0.5536x+R = 0.9741543棉子糖Km值y=0.5024x+底物：棉子糖R=21002468:(mol/mL)-1/v :mol/mLmin)-抑制的酶-酶促 力保存比例抑制的酶-酶促 力保存比

11、例酶上清液中酶活比率5.4-3-2-A_PY-SE:Incubation timeWhole cell enzyme 酶上清液中酶活比率5.4-3-2-A_PY-SE:Incubation timeWhole cell enzyme * Whole cell enzymeRatio of *.*123456789B_PY-Suc: Incubation time Catalyst activity Ratio of supernatent * Whole cell enzymeRatio of *.*123456789B_PY-Suc: Incubation time Catalyst act

12、ivity Ratio of supernatent 8765453200123456789B_培養(yǎng) 時間A_培養(yǎng) 時間 04809PY-PY-8765453200123456789B_培養(yǎng) 時間A_培養(yǎng) 時間 04809PY-PY-分析項酶酶PearsonSig.(2-Cross-N分析項酶酶PearsonSig.(2-Cross-NIncreasinglacticacid production rates,MinimizingIncreasinglacticacid production rates,Minimizinghigh glucose IncreasingcellReducing

13、 the toxic effectsfromsolvent aa生物膜形成a培養(yǎng)基營組細菌面樣貌bbbc*c*c*c*B_Crystal violet staining of floating OD570nm aa生物膜形成a培養(yǎng)基營組細菌面樣貌bbbc*c*c*c*B_Crystal violet staining of floating OD570nm 培養(yǎng)菌落總生物培養(yǎng)菌落總生物均5.4.1 細菌5.4.1 細菌衰老的延樣菌落總A發(fā)酵液加甘油B原始發(fā)酵樣菌落總A發(fā)酵液加甘油B原始發(fā)酵均a8PY-Suc-PY-7b6c5d*d*d*4cellcountsa8PY-Suc-PY-7b6c5d

14、*d*d*4cellcounts864-20-08處理 時間 64菌 落總 數(shù)（Log10 菌 落總 數(shù)唾液乳桿菌唾液乳桿菌2PY-PY-PY-SEPY-Suc864-20-08處理 時間 64菌 落總 數(shù)（Log10 菌 落總 數(shù)唾液乳桿菌唾液乳桿菌2PY-PY-PY-SEPY-Suc02468培養(yǎng) 時間 PY-Suc凍干PY-SE02468培養(yǎng) 時間 PY-Suc凍干PY-SE321（60：min;-0-05(A)60(B)50(酶保留比例4保留比例32(B)50(C)40 酶活酶活保留比例 酶活保留比例 保留比例 -酶保留比例321（60：min;-0-05(A)60(B)50(酶保留比

15、例4保留比例32(B)50(C)40 酶活酶活保留比例 酶活保留比例 保留比例 -酶保留比例 不同的菌體或細胞有著不同的 不同的菌體或細胞有著不同的標準大腸桿菌（Escherichiacoli）生物量達到釀酒酵母cerevisiae）動物細胞：107 乳酸菌乳酸菌高密度培養(yǎng)方細胞循環(huán)或截留實現(xiàn)乳酸菌高密度培固定化或微膠囊化高密度培養(yǎng)乳酸生物膜法高密度培養(yǎng)乳酸通過數(shù)學(xué)模型計算最佳培養(yǎng)條abN-N-1234567Different nitrogen Thenitrogenfrom1to7areYeastextract,Peptonefromfish,Bacterialpeptone,Biomass

16、 abN-N-1234567Different nitrogen Thenitrogenfrom1to7areYeastextract,Peptonefromfish,Bacterialpeptone,Biomass Catalyst pH6a54321bbcccccc0123456789Different carbon Thecarbonfrom1to9are:glucose,galactose,Catalyst pH6a54321bbcccccc0123456789Different carbon Thecarbonfrom1to9are:glucose,galactose,guarcatalyst activity pH.培養(yǎng) 時間 發(fā)酵液pH乙 酸鈉檸檬 酸鈉 磷酸氫 二鈉檸檬 酸銨 MRS基礎(chǔ)培養(yǎng) 基.培養(yǎng) 時間 發(fā)酵液pH乙 酸鈉檸檬 酸鈉 磷酸氫 二鈉檸檬 酸銨 MRS基礎(chǔ)培養(yǎng) 基PYSuc培養(yǎng)基白0.5%PYSuc培養(yǎng)基白0.5%0.5%，酵母粉蔗糖無水乙酸鈉0.6%,MgSO47H2OMnSO44H2O A:BiomassVsSEBiomassBiomass 2h 4h 6h 8h2h 4h 6h 8hA:BiomassVsSEBiomassBiomass 2h 4h 6h 8h2h 4h 6h 8h86B:CellcountsVsSECe