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由扩展青霉产生碱性脂肪酶的纯化及其特性 总被引:3,自引:0,他引:3
国外报道了不少由细菌、霉菌、酶母菌产生的中性脂肪酶及由细菌产生的碱性脂肪酶的纯化方法,国内尚未见有关脂肪酶纯化的报道。为了进一步研究和扩大该酶在医药、食品等方面的应用范围,我们对由扩展青霉突变株UN-503产生的碱性脂肪酶进行了纯化及其特性的研究,现报道如下。 相似文献
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脂肪酶是一类重要的工业用酶,广泛应用于诸多工业领域。与游离脂肪酶、物理或化学固定化脂肪酶相比,全细胞脂肪酶具有制备简单、无需蛋白质提取纯化、天然固定化、稳定性及抗逆性更好、制备及设备成本较低等优点,因此以全细胞形式利用脂肪酶被誉为是最有前景的降低生物转化成本的方法之一,关于全细胞脂肪酶的研究一直是脂肪酶领域的热点。就全细胞脂肪酶的研究进展进行归纳和述评,包括野生型全细胞脂肪酶和基因工程全细胞脂肪酶,并对其未来研究方向做出展望,以期为后续研究提供有益参考。 相似文献
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微生物脂肪酶的重组表达 总被引:1,自引:0,他引:1
微生物脂肪酶在传统和新型工业催化领域中的应用越来越广泛与深入。作为脂肪酶规模化制备主要途径的高效重组表达,为脂肪酶催化剂的最终形成及工业催化奠定了坚实的技术基础。概述并讨论了微生物脂肪酶重组表达的最新策略和发展趋势,阐述密码子优化、融合共表达、杂合启动子、同源高效表达、细胞表面展示和表达文库的高通量筛选等技术特点及其表达应用现状,指出细胞表面展示表达和表达文库高通量筛选体系为脂肪酶重组表达注入强劲活力;在此基础上对几种代表性微生物脂肪酶的重组表达进展作一综述,为微生物脂肪酶的重组表达研究及工业生产提供借鉴。 相似文献
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假单胞菌属脂肪酶的分子生物学研究进展 总被引:1,自引:0,他引:1
微生物脂肪酶是商品化脂肪酶的主要来源,并广泛应用于诸多工业领域。与其他微生物脂肪酶相比,细菌脂肪酶催化反应的类型更多、活性更高、稳定性更好,其中又以假单胞菌属(Pseudomonas)脂肪酶的性能最为优越。作为性能最为优越、应用最为广泛的一类脂肪酶,假单胞菌属脂肪酶研究一直是脂肪酶领域的热点。就假单胞菌属脂肪酶的分子生物学研究进展进行归纳和述评,包括基因资源挖掘及克隆、基因表达调控及分泌机制、活性过表达策略、蛋白质结晶及3D结构解析、蛋白质工程,并对其未来研究方向做出展望,以期为后续研究提供有益参考。 相似文献
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游离脂肪酶与固定化脂肪酶相比具有成本低、反应速率快等优势,是油脂化工中新的研究方向。前期研究表明,游离脂肪酶NS81006能高效催化多种油脂水解,进一步研究其对含独特羟基的绿色石油材料蓖麻油的水解过程,对于促进游离脂肪酶在新能源领域的应用具有重要意义。本文对影响游离脂肪酶NS81006催化蓖麻油水解过程的主要因素,温度、酶用量、水用量和搅拌速率进行了研究和优化,在优化后的条件下48 h水解率可达94.8%,且发现通过离心分离可有效实现NS81006的重复使用,连续回用5个批次,游离脂肪酶仍能有效催化水解反应。而对比高温高压法水解蓖麻油,发现游离脂肪酶NS81006具有明显优势。 相似文献
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Kei Arima Wen-Hsiung Liu Teruhiko Beppu 《Bioscience, biotechnology, and biochemistry》2013,77(11):1913-1917
In view point of the basic research for the enzyme properties and the development of utilization of lipase, a thermophilic fungus which could produce a remarkable amount of thermostable, alkalistable and extracellular lipase has been isolated from the compost soil. The taxonomical characteristics of this thermophilic fungus were examined and it was identified as Humicola lanuginosa S-38. 相似文献
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近年来溶胶-凝胶法固定脂肪酶已成为研究热点。选用TMOS、MTMS、ETMS和PTMS 4种硅烷试剂对黑曲霉脂肪酶进行了固定化研究。固定化的最佳配方为ETMS/TMOS=5:1、水与硅烷试剂分子比为8;固定化脂肪酶的固定率为80.2%、相对活性为136.3%;以乳化橄榄油作为底物,在50℃和pH4.0的条件下,固定化脂肪酶与游离脂肪酶Km分别为1.899×10-4M和2.789×10-4M;最适反应pH均为pH4.0,固定化脂肪酶在pH4.0~pH5.5之间其活性能保持95%以上;固定化脂肪酶最适反应温度为60℃,较游离脂肪酶提高了10℃;固定化脂肪酶的酸碱稳定性和热稳定性较非固定化酶有显著的提高。固定化脂肪酶的使用寿命和保存稳定性良好,使用12次后仍能够保留71.7%活性,在室温避光条件下保存180天后仍可保留79.2%活性。 相似文献
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Kinetics of mRNA and protein synthesis of genes controlled by the 1,4-beta-endoxylanase A promoter in controlled fermentations of Aspergillus awamori.
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R J Gouka H Stam A J Fellinger R J Muijsenberg A van de Wijngaard P J Punt W Musters C A van den Hondel 《Applied microbiology》1996,62(10):3646-3649
In this study, induction and repression kinetics of the expression of the Aspergillus awamori 1,4-beta-endoxylanase A (exlA) gene under defined physiological conditions was analyzed at the mRNA and the protein levels. Induction was analyzed by pulsing D-xylose to a sucrose-limited continuous culture of an A. awamori 1,4-beta-endoxylanase A (EXLA)-overproducing strain. Directly after the D-xylose pulse, exIA mRNA was synthesized, and it reached a constant maximal level after 45 to 60 min. This level was maintained as long as D-xylose was present. The kinetics of mRNA synthesis of the genes encoding Thermomyces lanuginosa lipase (lplA) and Escherichia coli beta-glucuronidase (uidA), which were also under the control of the exlA promoter, were similar to those observed for exlA mRNA. The repression of exlA expression was analyzed by pulsing D-glucose to a D-xylose-limited continuous culture. Immediately after the glucose pulse, the exlA mRNA level declined rapidly, with a half-life of approximately 20 to 30 min, and it reached a minimal level after 60 to 90 min. The time span between mRNA synthesis and the secretion of proteins was determined for EXLA and lipase. In both cases, mRNA became visible after approximately 7.5 min. After 1 h, both proteins became detectable in the medium but the rate of secretion of EXLA was faster than that of lipase. 相似文献
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Tomas Jacobsen Bo Jensen Jørgen Olsen Knud Allermann 《Applied microbiology and biotechnology》1989,32(3):256-261
Summary The imperfect fungus Geotrichum candidum produced extracellular lipase in a basic peptone-salt medium. By adding olive oil or Tween 80 to the basic medium the lipase yields could be enhanced and the maximal yields were found with Tween 80, which resulted in a sixfold increase in extracellular lipase activity as compared with basic medium. During the early phase of growth in medium with olive oil the proportion of cell-bound activity was higher than that of extracellular activity, and a delay in the secretion of extracellular lipase was found. The proportion of cell-bound activity from growth in basic medium and in basic medium with Tween 80 was lower than that of extracellular activity during the entire growth phase. Analyses by polyacrylamide gel electrophoresis showed that the lipase activity from growth in all three media could be ascribed to equivalent protein bands at 57 000 and 61 000 daltons. Immunodiffusion showed that the cell-bound preparation contained lipase that was immunologically identical with purified extracellular lipase from G. candidum. 相似文献
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【目的】克隆破囊壶菌Aurantiochytrium sp.PKU#SW7的脂肪酶基因,实现其在大肠杆菌细胞中异源高效表达,并进行初步酶学性质研究。【方法】基于转录组数据注释,获得脂肪酶基因lip,构建重组基因工程菌Rosetta(DE3)/p ET30-lip,利用双温控自诱导方法高效表达蛋白,表达产物(LIP)经Ni-Agarose His亲和层析柱纯化后进行酶学性质研究。【结果】从Aurantiochytrium sp.PKU#SW7中克隆得到一个大小为873 bp的脂肪酶基因(Gen Bank登录号为KT305964),该酶对p-NPB最适反应温度和pH分别为40°C和8.0。以不同浓度的金属离子Ca~(2+)和Co~(2+)溶液分别保温处理酶液30 min可使酶活提高1.3倍左右;甲醇对脂肪酶的抑制作用不明显。在最适反应条件下对p-NPP与p-NPB的酶活力分别为70.0±3.1 U/mg和102.5±2.6 U/mg。【结论】Aurantiochytrium sp.PKU#SW7脂肪酶具有良好的特性,符合生物柴油生物催化剂基本要求。 相似文献
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Inhibition of lipase activities by basic polysaccharide 总被引:1,自引:0,他引:1
Tsujita T Takaichi H Takaku T Sawai T Yoshida N Hiraki J 《Journal of lipid research》2007,48(2):358-365
Basic polysaccharide strongly inhibited the hydrolysis of trioleoylglycerol (TO) emulsified with phosphatidylcholine and taurocholate by either pancreatic lipase or carboxylester lipase. DEAE-Sephadex dose-dependently inhibited the hydrolysis of TO by pancreatic lipase and carboxylester lipase; however, carboxymethyl-Sephadex and Sephadex G-50 did not inhibit the hydrolysis. Polydextrose (PD), a soluble polysaccharide, was a very weak inhibitor of pancreatic lipase. However, when a basic group, a DEAE group, was attached to PD, lipase inhibition by DEAE-PD was increased, and this was dependent on the substitution ratio of DEAE groups. The number of positive charges per PD molecule is important in lipase inhibition. Similar substitution effects were observed with other basic groups, such as piperidinoethyl and 3-triethylamino-2-hydroxypropyl. The natural basic polysaccharide, chitosan, also inhibited pancreatic lipase activity. Gel-filtration experiments suggested that DEAE-PD did not bind strongly to pancreatic lipase. The effect of DEAE-PD on TO hydrolysis by pancreatic lipase was studied using various emulsifiers: DEAE-PD (50 microg/ml) did not inhibit the hydrolysis of TO emulsified with arabic gum, phosphatidylserine, or phosphatidic acid. In vivo, oral administration of DEAE-PD to rats reduced the peak plasma triacylglycerol concentration and increased fecal lipid excretion. These results suggest that basic polysaccharide is able to suppress dietary fat absorption from the small intestine by inhibiting pancreatic lipase activity. 相似文献
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【背景】脂肪酶是一类特殊的酯键水解酶,广泛应用于工业化生产中,微生物是工业脂肪酶的主要来源。瘤胃中微生物种类繁多、数量庞大,已有关于瘤胃微生物产纤维素酶的报道,尚无产脂肪酶瘤胃微生物的分离筛选报道。【目的】从牦牛瘤胃中分离筛选出能够产脂肪酶的微生物,并进行菌株鉴定及其酶学性质的研究。【方法】以橄榄油为唯一碳源,通过中性红油脂平板进行初步筛选后,用改进铜皂-分光光度法测定酶活力进行复筛;再经形态学观察、生理生化实验和16S rRNA基因序列分析进行菌种鉴定;研究3种脂肪酶的最适作用温度、pH值及金属离子、有机溶剂和表面活性剂对酶活力的影响。【结果】筛选出6株酶活力较高的菌株,其中3株为液化沙雷氏菌,2株为白地霉,1株为卷枝毛霉。脂肪酶的酶学性质研究表明:液化沙雷氏菌、白地霉和卷枝毛霉所产脂肪酶的最适作用温度为45、35和40°C;最适pH为8.0、7.0和7.0;Ca2+和Mg2+对3种脂肪酶均有激活作用;Zn2+对3种脂肪酶有不同程度的抑制作用,EDTA、SDS可使3种脂肪酶失活;3种脂肪酶对丙三醇的耐受力较高,卷枝毛霉脂肪酶对甲醇、乙醇、丙酮的耐受力较高。【结论】从牦牛瘤胃中分离出3种产脂肪酶的微生物,且证实瘤胃微生物在脂肪酶研究方面具有较高的价值。 相似文献
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果树核心种质研究进展 总被引:6,自引:0,他引:6
近年来,继果树种质资源收集保存和鉴定评价等重大项目相继开展之后,利用现有基本信息、鉴定评价数据构建果树核心种质已成为种质资源领域又一新的研究热点和发展方向。根据核心种质的内涵和果树自身特点,提出了果树核心种质研究的主要特性,阐述了其主要研究内容、方法和步骤等研究现状,并通过分析果树核心种质构建的特殊性,指出存在的主要问题,探讨了果树核心种质研究的重点和发展方向。 相似文献
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随着国家新农村建设和新型城市化的稳步推进,乡村景观格局的变化开始受到广泛关注,特别是相对闭塞的少数民族乡镇,理解其景观格局的时空变化对于探索民族乡镇发展至关重要.本研究基于景观生态学理论,利用典型达斡尔族少数民族乡镇——阿尔拉镇2008—2013年间的土地利用矢量数据,通过ArcGIS和 Fragstats软件计算景观格局指数,从类型水平和景观水平上对景观格局变化进行分析.结果表明: 在类型水平上,研究区农、林、草地逐渐向建筑用地转移;在景观水平上,景观多样性指数偏低,异质性较高,连通性不强,景观破碎化程度较高.将景观指数与社会经济指标进行典型性相关分析和灰色关联分析,发现人口的变化和GDP的增长是研究区景观格局变化的最主要驱动因子.最后,结合经济、文化、政策、自然等因素探讨了导致阿尔拉镇景观格局演变的主要驱动因素,并给出相应对策,该结果可为我国东北少数民族聚居区的发展提供基础信息和理论依据. 相似文献