产脂肪酶中度嗜盐菌ZF-03的分离鉴定与酶学性质研究

[目的]研究盐碱地产脂肪酶的中度嗜盐茵菌株的形态和生理生化特点及其所产胞外脂肪酶的性质。[方法]利用平板筛选法分离得到一株产脂肪酶中度嗜盐菌,对其进行形态、生理生化和分子水平的鉴定,并对其产生的脂肪酶的部分酶学性质进行研究。[结果]产脂肪酶的菌株命名为Nesterenkoniahalobiasp．ZF-03。对脂肪酶粗酶液酶学性质的研究结果显示该脂肪酶的最适反应温度在46～50℃之间,温度小于70℃时的稳定性较好;最适pH为8．5,在pH7．5～9．5范围内,相对酶活性较高,酶具有良好的稳定性;反应的最适NaCl浓度为0—3％,在0—5％NaCl浓度下有很好的稳定性。[结论]分离得到了一株产脂肪酶活性较高的中度嗜盐茵。     

“两广二号”家蚕抗病毒基因Bmlipase-1和BmSP-2克隆与原核表达

本研究以优良杂交品种“两广二号”家蚕为试材,克隆了该杂交品种家蚕两个抗家蚕核型多角体病毒（BmNPV）基因：脂肪酶基因Bmlipase-1和丝氨酸蛋白酶基因BmSP-2,测序并分别与不同品种蚕的同源基因序列进行比较。结果显示,“两广二号”家蚕Bmlipase-1基因ORF长度为885bp,编码294个氨基酸,BmSP-2扩增长度为855bp,编码284个氨基酸;它们的核苷酸和推导氨基酸序列同源性皆达92％以上,Bmlipase-1更保守,同源性大于99％;“两广二号”家蚕的Bmlipase-1基因脂肪酶活化部位和BmSP-2基因酶催化三联体位点的氨基酸残基与不同品种蚕的完全相同。以上结果说明这两个抗病毒基因在蚕的遗传进化过程中高度保守,提示其可能在机体消化或者免疫防御方面起着重要生理作用。将这两个抗病毒基因在大肠杆菌BL21中进行融合表达,获得的融合Bmlipase-1和BmSP-2蛋白分子量分别为47kD和42kD左右。     

Direct measurement of rice bran lipase activity for inactivation kinetics and storage stability prediction

Rice bran is a rich source of valuable nutrients and has potential for high-value applications. Endogenous lipases catalyze the hydrolysis of rice bran oil to free fatty acids, which initiates lipid oxidation. The evaluation of the success of rice bran stabilization processes in terms of the degree of lipid oxidation and shelf-life has so far relied on the measurement of free fatty acid content over a storage period of 3–6 months. In the present study, a photometric and a titrimetric pH-stat method for direct lipase activity measurement immediately after debranning were adapted to rice bran. The photometric method was further applied to determine rice bran lipase/esterase inactivation kinetics, which are useful to optimize stabilization treatments in order to prevent overprocessing and retain maximum level of nutrients. Rice bran was heat-treated in a specialized, hermetically sealable reactor at controlled holding times (5–40 min), temperatures (70–145 °C) and moisture contents (10–20%). Temperature dependency of the lipase/esterase inactivation rate could be described by the Arrhenius equation. Empirical findings on the importance of moisture content for effective rice bran stabilization could be quantified. Furthermore, the results demonstrate the great potential of the method to predict the shelf-life of stabilized rice bran without time-consuming storage tests.     

DM-130大孔树脂固定脂肪酶催化松香酯化反应的研究

采用DM-130大孔树脂为载体,对脂肪酶进行了固定化,利用所获得的固定化脂肪酶催化松香酯化,并由此获得了固定脂肪酶催化松香酯化的最佳反应条件:温度35℃,pH值6.0,加水量1%(体积比),分子筛量与反应液的质量比1∶40,最佳振荡反应时间36 h。     

超声波处理对鸭肉肌内磷脂酶解反应的影响

以脂质体为反应介质、鸭肉肌内磷脂水解酶为酶液,研究了超声温度、超声时间以及超声功率对鸭肉肌内磷脂酶解的影响。结果表明:随着超声温度的升高,磷脂酶解程度显著增加,但超过50℃时,磷脂酶解程度降低;随着超声时间的延长,磷脂酶解程度增加,在0.5 h内即可产生大量的酶解产物;随着超声功率的增大,磷脂酶解程度增加。鸭肉肌内磷脂酶解最优化的反应条件为:超声温度50℃、超声时间0.5 h、超声功率270 W。     

油菜种子含油量相关的脂肪酶基因BnSDP1克隆与表达分析

通过电子延伸和RACE (cDNA末端快速扩增) 克隆得到一个与油菜种子含油量相关的脂肪酶基因,命名为BnSDP1,其开放阅读框为2 472bp,编码分子量为92kD的蛋白,预测等电点为6.75,具有保守的patatin结构域,跨膜域位于N端和中间,序列分析显示BnSDP1拥有GXSXG脂肪酶的基本特征序列GSSVG。利用RT-PCR技术分析发现,在种子发育的成熟期（即脱水期）,BnSDP1在低油品系种子中的转录水平高于高油品系,在根、茎、叶、花中均有表达,但在根和叶片中表达量高,在根中尤其显著,且受蔗糖诱导上调表达。     

花生种子脂肪酶活力定量测定及其与储藏特性的相关性

对已有水稻脂肪酶活力比色测定方法进一步修改和优化,建立了花生脂肪酶活力定量测定方法,并确定取样量5 mg、底物浓度30 mg mL^–1为最适试验条件,该方法具有精确性好、特异性强、重复性好等特点。利用该方法检测216份花生种质资源的脂肪酶活性,表明不同材料间差异极显著(P<0.001)。试验筛选出酶活力高低差异显著的材料10份(高低各5份),并以其中2份高脂肪酶活性和2份低脂肪酶活性材料进行高温、高湿人工加速老化试验,结果表明,随老化时间延长,高脂肪酶活性材料发芽率下降较快,而低脂肪酶活性材料发芽率下降较慢,花生脂肪酶活力与其种子活力呈负相关(相关系数为–0.8875)。该结果为进一步深入研究脂肪酶对花生储藏劣变的影响提供了参考。     

Screening antiacne potency of Indonesian medicinal plants: antibacterial,lipase inhibition,and antioxidant activities

Indonesian medicinal plants were screened as potential sources of antiacne agents. The screening methods were performed using antibacterial assay against Propionibacterium acnes, lipase inhibitor assay, and antioxidant assay. The results showed that from 40 plant materials extracted with methanol and 50% ethanol in water, Caesalpinia sappan was the best extract based on the combined activities: antibacterial (minimum inhibitory concentration 0.13 mg/ml; minimum bactericidal concentration 0.25 mg/ml), lipase inhibitory [50% inhibitory concentration (IC₅₀) 120.0 μg/ml], and antioxidative (IC₅₀ 6.47 μg/ml). Another prospective extract is Intsia palembanica based on its lipase inhibitory activity (IC₅₀ 4.1 μg/ml) and antioxidant activity (IC₅₀ 3.87 μg/ml). Part of this report was presented at the 58th Annual Meeting of the Japan Wood Research Society, Tsukuba, March 2008