脂肪酶

The enzyme activity of LpsA2 was increased to 250% or above by 1 mmol/L Co2+, Hg2+ and Zn2+, respectively. The activity was also elevated to 110.7% and 138.0% by 15% dimethylformamide and dimethyl sulfoxide, and 352.7% and 189.7% by 0.1% and 1% Span-20, respectively. The enzymatic properties of LpsA2 from S. avermitilis were characterized that may provide the fundament to the screening of bio-engineered bacteria and the industrial applications in food processing and drug synthesis.

avermitilis的脂肪酶基因lpsA2进行了异源表达和酶学功能鉴定，不仅为脂肪酶的研究积累了更多数据，也为具有优良性能的脂肪酶生物工程菌的筛选奠定了基础，更为其在食品加工、药物合成等工业生产中的应用提供了依据。

The experiment two: enzyme preparation significantly improved average daily gainand feed conversion ratio (P＜0.05). Enzyme preparation significantly increased energymetabolizability and digestibility of crude fiber, crude protein and neutral detergent fiber,but had no remarkable effect on digestibility of dry matter, crude fat and acid detergentfiber. Enzyme preparation significantly decreased the relative viscosity of duodenal andjejunal digesta. The pH of intestine had no noticed difference in all groups. Enzymepreparation significantly decreased relative weight of gizzard, proventficulus, duodenum,jejunum and ileum. Enzyme preparation significantly increased villus size of duodenumand jejunum, and villus to crypt ratio of duodenum and ileum significantly increased too.Enzyme preparation considerably decreased ileal crypt height (P＜0.05), and didn"t affectthickness of intestinal wall. Supplementing enzyme preparation, the serum glucose, totalprotein and alanine aminotransferase, but enzyme preparation hadn"t noticed influenceupon uric acid, total cholesterol, triglyceride and high-density lipoproteins. Enzymepreparation significantly increased insulin, triiodothyronine and insulin-like growthfactor-Ⅰ. Adding enzyme preparation, the percentage of thyroid stimulating hormone andgrowth hormone in the serum increased 16.44%, 19.18% and 18.84%, 21.74%respectively, and the percentage of glucagon and thyroxine decreased 12.07%, 14.36% and 13.79%, 15.40%, but failed to reach statistical significance (P＞0.05). Enzymepreparation significantly increased (P＜0.05) the trypsin and amylase activity of duodenaland jejunal digesta, but enzyme preparation didnt affect significantly (P＞0.05) theintestinal lipase activity and pancreatic digestive enzyme. Enzyme preparation had nosignificant effect on caecal microbial population.

试验二：酶制剂显著提高平均日增重和饲料转化率(P＜0.05)；酶制剂显著提高能量代谢率及粗纤维、粗蛋白、中性洗涤纤维消化率(P＜0.05)，而对干物质、粗脂肪、酸性洗涤纤维消化率影响不显著；酶制剂显著降低十二指肠和空肠食糜相对粘度(P＜0.05)；添加酶制剂对肠道pH影响不显著；酶制剂显著降低肌胃、腺胃、十二指肠、空肠、回肠相对重(P＜0.05)，显著提高十二指肠和空肠绒毛高度，显著增加十二指肠和回肠绒毛高度／隐窝深度，降低回肠隐窝深度(P＜0.05)，对肠壁厚度影响不显著；酶制剂显著提高血清葡萄糖、总蛋白和谷丙转氨酶浓度(P＜0.05)，对尿酸、总胆固醇、甘油三酯及高密度脂蛋白浓度影响不显著，显著提高胰岛素、T_3、IGF-Ⅰ水平，添加酶制剂后，促甲状腺激素、生长激素分别提高16.44％、19.18％和18.84％、21.74％，胰高血糖素和T_4分别降低12.07％、14.36％和13.79％、15.40％，但差异不显著；酶制剂对胰腺消化酶活性影响不显著，显著增加十二指肠和空肠胰蛋白酶、淀粉酶活性，对小肠脂肪酶活性影响不显著；酶制剂对盲肠微生物菌落数影响不显著。

The different factors involved in the process of the folding and activation of the microbial lipase are reviewed in this article,including lipase-specific foldase,lipase activation factor,prosequence,calcium ion and disulphide bond.

对微生物脂肪酶活性构像的形成和激活机制进行深入研究，有助于我们更好地了解脂肪酶的结构和功能，为脂肪酶基因的大量表达和体外定向进化奠定基础。本文主要综述了作为脂肪酶结构成分的多种因子在脂肪酶活性构像形成、激活及稳定中的作用。具体催化反应体

Signal peptide sequence was removed lest it couldnt be recognized by Bacillus subtilis, gene coding for mature peptide was ligated to downstream of sacB signal peptide sequence of pWB980 to form a new ORF and generate pWB980-LipA, gene coding for its chaperone was also amplified and ligated to the downstream of LipA to generate a secretion/expression vector pWB980-LipAB.thechaperone gene was sequenced and analyzed by multiple alignments, resulted showed that there were mutations of nucleotides(1～7) and amino acids (1～2) with the other reported genes.

为了防止枯草芽孢杆菌信号肽酶不能识别脂肪酶的信号肽，切除掉脂肪酶的信号肽编码序列，与枯草芽孢杆菌分泌表达质粒pWB980连接并与质粒上的SacB信号肽构成一个完整的开放式阅读框，获得重组质粒pWB980-LipA，因为该脂肪酶的活性表达需要一个特异性分子伴侣帮助折叠成有活性的构象，将脂肪酶分子伴侣基因串连到脂肪酶基因的下游，获得重组分泌表达载体pWB980-LipAB并且测序分析分子伴侣的基因序列和氨基酸序列并与其它报导的序列进行了比对，发现有1到7个碱基的差异和1到2个氨基酸残基的突变。

In addition, SW480 and HCT116 cells were treated with deoxycholic acid for 15, 30, 45, 60, 90 and 120 minutes. It was found that the deoxycholic acid treated cells showed an elevation of ATP activity by 12.35 %, 20.08 %, 29.69 %, 33.56 %, 45.85 %, 54.32 %, respectively. While the HCT116 treated cells, showed an elevation of ATP activity by 3.69 %, 5.59 %, 8.52 %, 12.26 %, 16.52 %, 19.71%, respectively.From the experiments conducted using the cell lines described above, it was further confirmed that genes CYP7A1 (cytochrome P450, family 7, subfamily A, polypeptide 1), AKR1D1 (aldo-keto reductase family 1, member D1), ALDH1A1 (aldehyde dehydrogenase 1 family, member A1), BAAT (bile acid Coenzyme A: amino acid N-acyltransferase), ABCA1 (ATP-binding cassette, sub-family A, member 1), APOC3 (apolipoprotein C-III), LIPA, LIPE (lipase, hormone-sensitive), PTGER2 (prostaglandin E receptor 2) and PTGS2 (prostaglandin-endoperoxide synthase 2) were up-regulated.

此外，为了证实其脂肪代谢产物胆酸对细胞癌化的影响，亦使用SW480和HCT116细胞株，经由脱氧胆酸在15分钟、30分钟、45分钟、60分钟、90分钟和120分钟处理后，其细胞增生比率分别有12.35%，20.08%，29.69%，33.56%，45.85%和54.32%(p.05)以及3.69%，5.59%，8.52%，12.26%，16.52%和19.71%(p.05)上升情形，并从这些细胞株进一步证实CYP7A1(细胞色素P450-7A1)、AKR1D1(醇醛酮还原酶-1-D1)、ALDH1A1(醛脱氢酶-1-A1)、BAAT(胆酸辅酶A：氨基酸N-醯基转移酶)、ABCA1(三磷酸腺苷结合盒转运体A1)、APOC3、LIPA、LIPE(脂肪酶/激素敏感脂肪酶)、PTGER2(前列腺素E受体2)和PTGS2(前列腺素内过氧化物合成酶2)等基因表现皆有呈现向上调控的情形。

Lactobacillus plantarum had lipolytic activity. Bacillus subtilis not only had the lipolysis but also had proteolysis.Single strains fermenting experiment showed that, the effect of different strain to the pH, acidity, the total amino acid were not signifient, Pediococcus pentosaceusand Lactobacillus plantarum could improve the content of free amino acid in dried beef.

研究结果表明：沃氏葡萄球菌、腐生葡萄球菌和植物乳杆菌只有脂肪酶活性，无蛋白酶活性；松鼠葡萄球具有弱蛋白酶活性；木糖葡萄球菌具有弱脂肪酶活性；戊糖片球菌具有脂肪酶活性，而蛋白酶活性较弱；枯草芽孢杆菌具有两种酶活性。

In this study, isolate 23 microbes with lipase activity from Wu-Jie scrap heap at Ilan and Taichung Environmental Protection Section. When the microbes incubator in 50oC, there are isolates B61 and M63 that lipase activity are better than others. Brevibacillus borstelensis SH168 isolated from NTU resterant food waste compost is used as control check. Isolates B61, M63 and Brevibacillus borstelensis SH168 determine the lipase activity by the LB with tributyrin. The lipase activity of isolates B61 and M63 are 2~3 fold higher than Brevibacillus borstelensis SH168. So isolates B61 and M63 are more useful in producing lipase. Isolate B61 can grow at pH 5~9, and at temperature 20oC ~ 60oC; isolate M63 can grow at pH 6~10, and at temperature 30oC ~ 50oC. Isolates B61 and M63 were identified by 16S rRNA. Isolate B61 is Bacillus circulans, and isolate M63 is Bacillus species.

本研究，於宜兰五结垃圾场和台中市环保局厨余堆肥样品中共分离出 23 株具脂肪酶活性菌株， 23 株菌株中经过三油酸甘油酯的洋菜培养基於 50℃培养 5 天后，发现有 2 株菌的脂肪酶活性较强，分别为分离株 B61 和 M63；以本实验室先前自台大女九餐厅蔬果厨余堆肥中所筛选出的耐高温脂解菌 Brevibacillus borstelensis SH168作为本实验对照菌株，将分离株 B61、M63 及 Brevibacillus borstelensis SH168 3 株菌株於含有三酪酸甘油酯液态培养基培养并测定其脂肪酶活性，发现分离菌株之活性皆比 Brevibacillus borstelensis SH168 高出 2~3 倍，认为分离株 B61、M63 适於产生脂肪酶的菌株，分离株 B61 在 pH 5~9 皆可生长，生长温度介於 20 oC ~ 60oC；分离株 M63 在 pH 6~10 可生长，生长温度介於 30 oC ~ 50oC； 2 株分离菌株以 16S rRNA定序结果分离株 B61 为 Bacillus circulans，分离株 M63 为 Bacillus 属。

I developed bee protein feed high in protein, fats, minerals, fat soluble vitamins, the proportion of suitable water-soluble vitamins, feed ratio of computational science, in line with bee nutritional requirements; feed is also appropriate to add grape oxidase, amylase, invertase, protein hydrolytic enzymes, fat bees enzymes necessary enzymes, raising bees on nutrient absorption rate, while reducing excretion, so that more fully reflects the use of nutrition.

我开发蜂蛋白饲料高蛋白质，脂肪，矿物质，脂肪和水的比例适当的水溶性维生素，饲料比例的计算科学，与蜜蜂营养要求，饲料也应加重葡萄氧化酶，淀粉酶脂溶性维生素，蔗糖酶，蛋白水解酶，脂肪酶的必要蜜蜂酶，提高对养分的吸收率蜜蜂，同时减少排泄，因此更充分地反映了营养的使用。

The protease activity in foregut was higher than that of midgut in an acidic condition at pH 5.0-5.8, while in an alkalescence condition at pH 7.0-8.6, the protease activity was higher in midgut than that in foregut. The lipase activity of foregut was high and stable in two pH ranges: 4.2-5.0 and 6.2-7.0, and that of midgut was high at pH 3.8, while the lipase lost enzyme activity at pH over 9.0. The amylase activity was high and stable at pH 6.6-7.4 in both foregut and midgut. The cellulase activity of foregut and midgut was high and stable at pH 6.2-7.4 and 5.4-7.0 respectively.

刺参前肠蛋白酶活力在酸性环境下较中肠高，且在反应pH为5.0-5.8之间酶活力相对稳定，而中肠蛋白酶活力在碱性环境下较前肠高，且在反应pH为7.0-8.6之间酶活力相对稳定；刺参前肠脂肪酶活力随着pH的升高出现两个相对稳定的峰值，分别为4.2-5.0和6.2-7.0区间，中肠脂肪酶活力在pH为3.8时达到最大值，而在pH超过9.0明显失活；刺参前肠和中肠淀粉酶活力在pH变化时表现出相似的变化趋势，在反应pH为6.6-7.4之间酶活力较高且相对稳定；刺参前肠和中肠中纤维素酶活力在pH变化时反应不一致，前肠淀粉酶在反应pH为6.2-7.4之间酶活力较高且相对稳定，中肠纤维素酶活力在反应pH为5.4-7.0之间酶活力较高且相对稳定。

Compared the activities of 5 sorts of lipases from different source in synthesis of fatty acid esters and hydrolysis of olive oil, 3 sorts of lipases are better. Compared the stability and the activities of 3 sorts of Upases in different media, organic solvents and CAL lipase are best.

通过对不同来源的5种脂肪酶分解橄榄油和合成脂肪酸酯的比较，选出了合成脂肪酸酯活力较高的3种脂肪酶，通过比较不同介质中这3种脂肪酶的合成活力后，得出这三种酶在有机相中活力最高，从操作稳定性角度出发，选出催化酯合成效果最好的脂肪酶CAL。

According to the clear water experiment, aeration performance of the new equipment is good with high total oxygen transfer coefficient and oxygen utilization ratio.

曝气设备的动力效率在叶轮转速为120rpm～150rpm时取得最大值，此时氧利用率和充氧能力也具有较高值。

The environmental stability of that world - including its crushing pressures and icy darkness - means that some of its most famous inhabitants have survived for eons as evolutionary throwbacks, their bodies undergoing little change.

稳定的海底环境─包括能把人压扁的压力和冰冷的黑暗─意谓海底某些最知名的栖居生物已以演化返祖的样态活了万世，形体几无变化。