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1.
广东经济鱼类不饱和脂肪酸的含量分析与组成特征评价   总被引:1,自引:0,他引:1  
张文凤  黄伟雄 《环境化学》2020,39(5):1181-1191
采用水解提取-气相色谱法测定鱼肉中脂肪酸的含量和组成,采用元素分析仪-同位素比值质谱法(EA-IRMS)测定鱼肉碳稳定同位素比值(δ~(13)C).开展广东沿海地区主要经济鱼类肌肉脂肪酸的含量分析及组成特征评价研究.研究表明,25种经济鱼类鱼肉中主要含有单不饱和脂肪酸、n-6和n-3系列多不饱和脂肪酸,以及少量反式脂肪酸(TFAs);鱼肉的优势脂肪酸为油酸(OA)、亚油酸(LA)和二十二碳六烯酸(DHA);鲻鱼含有较高的二十碳五烯酸(EPA)和DHA,合计370.0 mg·(100 g)~(-1);鲻鱼鱼肉中TFAs的含量(280.0 mg·(100 g)~(-1))远高于其它品种鱼类;鲢鱼和黑鲷EPA+DHA的含量较高,分别为(154.4±100.0 mg·(100 g)~(-1)和1287±291.1 mg·(100 g)~(-1),两种鱼均具有较好的商业开发价值;统计学分析结果表明了鱼类的不饱和脂肪酸在组成上具有种属差异,揭示了浮游食性的鱼类其鱼肉组织中MUFAs、n-6 PUFAs以及n-3 PUFAs的含量高于底栖食性的鱼类.鱼肉脂肪酸的组成差异较大,表现为品种、空间以及食性差异.建议市民在日常膳食中丰富食鱼的种类和数量,警惕过多地食用单一品种鱼类(尤其是鲻鱼)而导致TFAs摄入过多的风险,从而确保鱼类脂肪酸的平衡摄入.  相似文献   

2.
利用重叠延伸PCR方法对扩展青霉脂肪酶基因进行体外定点突变,构建了K115R与随机突变体ep8双突变的重组质粒pAO815-ep8-K115R.将该质粒电转化到毕赤酵母GS115中进行异源表达,获得双突变体脂肪酶PEL-ep8-K115R-GS.与野生型脂肪酶PEL-GS和随机突变体脂肪酶PEL-ep8-GS进行比较,发现双突变体脂肪酶PEL-ep8-K115R-GS在40℃温浴30 min后残余酶活为89%,比野生型脂肪酶PEL-GS和随机突变体脂肪酶PEL-ep8-GS分别提高54%和27%;双突变体脂肪酶PEL-ep8-K115R-GS的Tm值为42.2℃,比野生型脂肪酶PEL-GS提高3.5℃,比随机突变脂肪酶PEL-ep8-GS提高2.0℃.研究结果表明对扩展青霉脂肪酶的分子改造能提高其热稳定性.  相似文献   

3.
生物柴油是一种可再生且环保的化石燃料的替代品,研究固定化酶生产生物柴油对解决石化燃料耗尽和环境保护具有重要意义.将制备的纳米材料Fe_3O_4@SiO_2-CHO与伯克霍尔德氏菌脂肪酶以共价交联的方式固定,通过响应面法(RSM)优化其固定化的条件,然后以固定化酶催化橄榄油生产生物柴油,并对影响生物柴油制备的因素进行优化,最后通过GC-MS检测转酯产物的成分.结果表明:在加酶量30.22 m L、戊二醛浓度2%、固定化温度40℃、固定化时间4 h时得到最大的固定化脂肪酶酶活和固定化效率,分别为10 038 U/g和96.9%.然后以此最佳条件制得的固定化酶催化制备生物柴油,得到最优条件为加酶量1.5 g、醇油比4:1、反应温度30℃、反应时间4 h.最后用GC-MS分析所得产物,得到最大转酯效率为98.84%,且固定化酶重复转酯10次后,转酯效率仍可达90%.本研究获得的Fe_3O_4@SiO_2-CHO固定化酶不仅能在短时间内催化得到很高的生物柴油产量,且易回收、可重复使用、生产成本大大降低,是工业化生产生物柴油的一种高效环保的生物酶催化剂.  相似文献   

4.
白地霉(Geotrichum candidum)经过发酵培养,以发酵液为lip42脂肪酶粗酶液,经硫酸铵分级沉淀、透析脱盐、DEAE纤维素柱层析及Sephadex-75凝胶柱层析纯化处理,脂肪酶比活力达到121.68U/mg,纯化倍数为粗酶液的7.3倍,回收率为30.25%,经SDS-PAGE电泳显示为单一条带,相对分子质量约64×103.脂肪酶的酶学性质研究结果表明:在40℃以下温度保存1h内酶活性相对稳定;在pH8.0和30℃反应条件下表现相对高的酶活性;在Mg2+存在情况下酶活性提高10%左右,而其它离子如Cu2+、Zn2+等以及0.1%的Tween80、Tween20及Triton-100对酶活性却有不同程度的抑制作用,1mmol/L的SDS和EDTA-Na对酶活性的影响很微小.  相似文献   

5.
高产脂肪酶菌株的筛选鉴定及酶学、转酯特性   总被引:2,自引:0,他引:2  
从自然环境中筛选水解酶活高且酶学、转酯特性优良的产脂肪酶菌株,对脂肪酶工业化发酵生产及生物柴油制备的研究具有重要意义.采用罗丹明B平板初筛和摇瓶发酵复筛法,从70份含油脂丰富的样品中筛选产脂肪酶酶活较高的菌株进行16S rRNA鉴定,研究其酶学性质;用大孔树脂固定酶,在无溶剂体系中催化橄榄油制备生物柴油,研究其转酯特性.结果筛选到一株高产脂肪酶的菌株WZ10-3,通过p-NPP法测得其初始酶活为78.68 U/mL,经16S rRNA鉴定属于伯克霍尔德氏菌(Burkholderia sp.),与B.stabilis同源性达到99%.该菌在发酵48 h时达到产酶高峰,所产脂肪酶的最适作用温度为50℃,最适作用pH为7.0,70℃下的半衰期可达1 h,pH为7-9时稳定性良好.以大孔树脂NKA-9和HPD600为载体制备的2种固定化脂肪酶,催化橄榄油生产生物柴油的转酯率均可达到97%.综合表明,菌株WZ10-3脂肪酶的初始水解酶活高于大多数野生脂肪酶,热稳定性好且转酯特性优良,有很好的后续研究价值.图7表3参25  相似文献   

6.
响应面法优化海洋细菌MP-2酯酶发酵条件   总被引:4,自引:1,他引:3  
采用响应面法(Response surfsce methodology)对海洋细菌MP-2酯酶的发酵条件进行了优化.首先采用了Placken-Burman方法对影响酯酶发酵的因素进行评价,筛选出了3个显著因素为豆饼粉、花生粕和种龄,并利用DesignExpert 7.0.3软件完成RSM法对显著性因素的进一步优化从而确定最佳条件.当花生粕0.9%;豆饼粉2.29%;接种量5·98%时,此时预测的最大酯酶活力为334.46 U/mL.在最佳发酵条件下,优化后该酯酶活力由258.8 U/mL提高到318.2 U/mL.实际酶活力达到理论预测值的95%,十分接近.  相似文献   

7.
从厌氧发酵污泥中筛选到一组高效、传代稳定的厌氧纤维素降解复合菌系L-3.该复合菌系的内切葡聚糖酶活(Cx)、滤纸酶活(FPA)、外切葡聚糖酶活(C1)、β-葡聚糖苷酶活(β-glucodase)分别为0.216、0.101、0.132、0.002U/mL;该复合菌系可使滤纸在42h内溃烂,并能在降解纤维素的同时产生氢气,气体中氢气含量最高可达70.2%,d13时滤纸失重率为70.6%.DGGE结果表明,该复合菌系主要由14种菌组成.在所选实验条件内,该复合菌系产纤维素降解酶的最适条件为:最佳碳源为滤纸,最佳氮源为硫酸铵,温度36℃,pH6.5~7.0,接种量5%.  相似文献   

8.
为开发适应性好的工业用酶,从深圳福田红树林自然保护区土壤中分离纯化获得一株产耐高温嗜碱性酯酶的菌株,通过形态学和分子生物学初步鉴定该菌株为芽孢杆菌属(Bacillus sp.),命名为Bacillus sp.W77;对所产的粗酶液进行酶活测定,发现该菌产的酯酶的最适反应温度是50℃,最适pH为8.0,且在碱性条件下较稳定;采用响应面法(Response surface methodology)对土壤菌株Bacillus sp.W77的产酯酶条件进行了优化,酵母提取物6.67 g/L,磷酸氢二钾0.56 g/L,硫酸镁0.36 g/L时,此时预测的最大酯酶活力的D405 nm值为1.266,在最佳产酶条件下,优化后酯酶活力的D405 nm值由0.829提高到1.235,实际值达到理论预测值的97.5%.  相似文献   

9.
从四川海螺沟原始森林腐土中分离到一株嗜酸性产纤维素酶的真菌X-13,其主要特点是产纤维素酶的最适pH及其纤维素酶最适反应pH均为2.0.在PDA培养基上培养时菌落呈浅黄色至肉桂色,反面呈黄色至棕褐色,产黄色色素;菌丝体透明有隔膜,分生孢子呈球形或近球形.根据菌株的形态特征以及ITS序列同源性和系统发育分析结果,鉴定该菌株为土曲霉(Aspergillus terreus Thom).该菌最佳产酶培养时间为8~10 d;最适产酶温度为30℃,纤维素酶最适反应温度为50℃;最佳碳源、氮源分别为纤维素粉和硫酸铵.通过响应面法对菌株产纤维素酶条件进行优化,使菌株X-13纤维素酶活从1.39 IU/mL提高到2.94 IU/mL,提高了111.5%.  相似文献   

10.
采用DNS法研究了我国广泛分布的一种低等木食性白蚁——黑胸散白蚁纤维素酶的体外酶活特性以了解其纤维素降解机制.结果表明,内切β-1,4-葡聚糖酶(EG)、纤维二糖水解酶(CBH)和β-葡萄糖苷酶(BG)这3种酶的最佳反应时间均为15 min,最佳底物浓度为1%,最适反应pH为5.6,最适反应温度为35℃.在最适反应条件下,EG、CBH和BG的活性分别达到71.3(±13.9)U/mg、5.8(±0.8)U/mg和4.1(±0.7)U/mg.EG在体外的热稳定性较差,在50℃及更高温度酶活很低或完全失活,但该酶对pH稳定性较好,在pH 3.2~8.0范围内酶活力变化不大.Native-PAGE电泳检测到该白蚁体内至少有8种不同的EG活性条带,肠道不同部位纤维素酶活性条带种类不同.这些研究表明,木食性白蚁降解纤维素是一个复杂的过程,需要多种纤维素酶的共同作用.  相似文献   

11.
To test whether heterotrophic protists modify precursors of long chain n−3 polyunsaturated fatty acids (LCn−3PUFAs) present in the algae they eat, two algae with different fatty acid contents (Rhodomonas salina and Dunaliella tertiolecta) were fed to the heterotrophic protists Oxyrrhis marina Dujardin and Gyrodinium dominans Hulbert. These experiments were conducted in August 2004. Both predators and prey were analyzed for fatty acid composition. To further test the effects of trophic upgrading, the calanoid copepod Acartia tonsa Dana was fed R. salina, D. tertiolecta, or O. marina that had been growing on D. tertiolecta (OM-DT) in March 2005. Our results show that trophic upgrading was species-specific. The presence of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the heterotrophic protists despite the lack of these fatty acids in the algal prey suggests that protists have the ability to elongate and desaturate 18:3 (n−3), a precursor of LCn−3PUFAs, to EPA and/or DHA. A lower content of these fatty acids was detected in protists that were fed good-quality algae. Feeding experiments with A. tonsa showed that copepods fed D. tertiolecta had a significantly lower content of EPA and DHA than those fed OM-DT. The concentration of EPA was low on both diets, while DHA content was highest in A. tonsa fed R. salina and OM-DT. These results suggest that O. marina was able to trophically upgrade the nutritional quality of the poor-quality alga, and efficiently supplied DHA to the next trophic level. The low amount of EPA in A. tonsa suggests EPA may be catabolized by the copepod.  相似文献   

12.
During early development, oviparous fish species must use finite lipid and fatty acid (FA) reserves for both catabolism and structural components. In cold environments, developing fish have the additional constraint of maintaining membrane fluidity for metabolic efficiency (homeoviscous adaptation), resulting in further demand on lower melting point FAs like n-3 polyunsaturated fatty acids (PUFAs). To examine whether marine fish embryos physiologically adapt to changing temperature environments, we incubated Pacific cod (Gadus macrocephalus) eggs at 5 temperatures (0, 2, 4, 6, and 8 °C) in the laboratory and sampled them repeatedly during development to measure changes in lipid/FA composition. Pacific cod embryos increased n-3 PUFA content during the egg stage in all temperature treatments, with the possible exception of 0 °C, where poor survival and hatch success limited our ability for continued sampling. At the beginning of the hatch cycle, free-swimming embryos shifted from lipogenesis to lipid catabolism. The rates of lipogenesis and catabolism were temperature dependent, and the distinct increase in unsaturated fatty acids at temperatures <8 °C was consistent with homeoviscous adaptation theory. However, with the possible exception of embryos at 0 °C, the relative amounts of essential fatty acids (e.g., EPA, DHA, AA) were conserved in a similar manner across incubation temperatures. Collectively, these data suggest Pacific cod are capable of homeoviscous adaptation but cannot tolerate temperatures approaching 0 °C despite their possible ability to biosynthesize PUFAs from other energetic sources.  相似文献   

13.
二十二碳六烯酸(Docosahexaenoic acid,DHA C22:6n-3)是具有各种重要生理功能的高度不饱和脂肪酸.分别以质粒pYTFD5和pYFAD4为模板,扩增获得860 bp的△5-延长酶基因(elo5)和1 600 bp的△4-脱饱和酶基因(fad4).利用重叠延伸PCR构建elo5-fad4融合基因,Hind Ⅲ/Sph Ⅰ双酶切后连接到经同样处理过的pYES2.0载体,构建重组表达质粒pYEL05-FAD4.转化酿酒酵母尿嘧啶缺陷型菌株INVScl,通过缺少尿嘧啶的选择性培养基筛选阳性克隆子.添加外源脂肪酸C20:5底物.半乳糖诱导表达.气相色谱-质谱分析表明,重组酵母总脂肪酸中出现了DHA(二十二碳六烯酸,C22:6n-63)新产物,融合基因elo5-fad4在酿酒酵母中得到了表达.图7表1参18  相似文献   

14.
Fatty acid biomarkers and stable isotope signatures were used to identify the sources of particulate and sedimentary organic matter and its input into the food web through the dominant consumer within the mangrove-dominated Rufiji estuary, Tanzania. Specific fatty acids were used to identify the preferred basal sources of dominant fauna (i.e. filter feeder bivalves, snails, crabs, shrimps, and three fish species), and their presence in the water and sediment samples in the estuary. Both fatty acid and stable isotope results revealed that food web in the Rufiji estuary depended on a variety of carbon sources (mangroves, allochthonous terrestrial inputs, macroalgae, and phytoplankton), contributing to a different degree into the diets of primary consumers and members of near-shore fish, but none of them were obligatory for the survival of these species. The δ15N values of major primary producers and consumers/predators revealed a trend for δ15N enrichment with increasing trophic level. The ratio of docosahexaenoic acid to eicosapentaenoic acid (DHA:EPA) decreased from pelagic to benthic feeding fish. This indicated that fish with different feeding modes derived their fatty acids from different primary sources of nutrition, and suggested that the DHA:EPA ratio may be a useful indicator of feeding mode.  相似文献   

15.
The distribution of n-3 highly unsaturated fatty acids (HUFA) over the major neutral and polar lipid classes was determined for two predominant types of live food used in the larviculture of marine fish and shrimp, i.e. freshly hatched and HUFA-enriched Artemia, and compared with data reported in the literature for wild copepods, representing the natural diet of these larvae. Lipid class composition and their content of n-3 HUFA, particularly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), were assessed in freshly hatched, HUFA-enriched and subsequently starved Artemia franciscana. The n-3 HUFA enrichment was based on feeding Artemia a lipid emulsion in which either fatty acid ethyl esters (EE, diluted with olive oil) or triacylglycerol (TAG) provided a level of 30% n-3 HUFA. Enrichment of Artemia with either type of the lipid emulsions resulted in an increase of total lipid content from 20.0 to 28.2–28.7% of dry matter mainly due to the accumulation of neutral lipid, primarily TAG (from 82 to 158 mg g−1 dry wt in freshly hatched and 24-h enriched Artemia). Enriched brine shrimp utilized up to 27–30% of their TAG content during 72 h of starvation at 12 °C. The absolute tissue concentrations of polar lipids remained constant at 71 to 79 mg g−1 dry wt throughout the enrichment and subsequent starvation. The level of n-3 HUFA increased drastically during enrichment from 6.3% of total fatty acids (8.2 mg g−1 dry wt) in freshly hatched nauplii to between 20.4 and 21.8% (40.4 to 43.2 mg g−1 dry wt) in 24-h enriched Artemia and was not significantly affected by the source of n-3 HUFA. During starvation, 18:0, 20:4n-6 and 20:5n-3 were retained, whereas 18:4n-3, 22:5n-3 and 22:6n-3 were specifically catabolized. The major polar lipids, phosphatidylethanolamine (PE) and phosphatidylcholine (PC), of freshly hatched Artemia showed very low levels of DHA (<0.1% of total fatty acids) and carried about 45% of the total EPA present. Enrichment with either of the emulsions resulted in an increase of the neutral lipid fraction which concentrated >64% of the EPA and >91% of the total DHA present. This is in sharp contrast with the high levels of n-3 HUFA, in particular DHA, in the polar lipid fraction reported for wild copepods. The contrasting distribution of DHA in the neutral and polar lipid fractions of enriched brine shrimp compared to the natural diet may influence the efficacy of this essential fatty acid for marine fish larvae in aquaculture systems. Received: 10 June 1997 / Accepted: 8 August 1997  相似文献   

16.
A 6-week feeding trial was conducted with 44-d-old European sea bass (Dicentrarchus labrax L.) in order to examine the effect of various dietary phospholipid (PL) sources on the incorporation of n-3 highly unsaturated fatty acids (HUFA) in tissue lipids. From weaning onwards the fish received diets prepared by coating different lipid fractions (7.5% diet) on an extruded basal diet (92.5% diet). The two PL-free control diets contained 0.5 and 2% of an emulsifier blend, respectively. Seven other diets contained 2% PL, differing by their purity and origin (vegetable or animal). All diets were rendered isolipidic by the addition of hydrogenated coconut oil. Feeding the PL-supplemented diets, except the diet containing hydrolyzed soybean PL (lyso PL), resulted in a higher survival and a 10 to 30% better growth as compared to the PL-free diets. No difference according to the PL origin was observed. The sea bass final lipid content increased with increasing body weight. Also the lipid class composition of the fish was clearly correlated with the final weight gain. Total neutral lipid increased from 51% of total lipid (initial fish) to 76% for fish fed the PL-free diets, and up to 88% for fish fed the sunflower PL. Weaning the fish on the experimental diets induced important changes in their fatty acid profiles characterized by a decrease in 18:3n-3, 20:5n-3 and 20:4n-6 and an increase in saturated fatty acids and 22:6n-3 (DHA). According to the fatty acid composition of both total and polar lipid, the weaned fish could be divided into three groups reflecting the dietary fatty acids: a group fed the vegetable PL, a group fed the animal PL and a PL-deprived group. An effect of dietary PL on the incorporation of dietary n-3 HUFA, more particularly DHA, was noticed. For a similar supply of DHA through the neutral lipids in the diet, fish fed PL-supplemented diets (except for the lyso PL diet) had 10 to 25% higher DHA levels in total and polar lipid than PL-deprived fish. This PL effect was already clear at the end of the weaning and was not related to the presence of n-3 HUFA in the PL source, as suspected in a previous study when feeding egg yolk PL. A better incorporation of DHA was not obtained by replacing the PL by an emulsifier or by lyso PL with higher emulsifying properties. Present results confirm a role of dietary PL in the absorption of dietary neutral lipids, by a mechanism other than emulsification. Received: 27 May 1997 / Accepted: 30 June 1997  相似文献   

17.
采用松花粉垂钓法分离到一株Docosahexaenoicacid(DHA)高产菌FJU 512.该菌株DHA含量高(占总脂肪酸的56. 24 % ),其它长链杂酸含量少(仅有docosapentaenoicacid, DPA),极具开发应用价值.高密度培养可获得33gL-1生物量.该菌株行二分裂生长,没有分生胞子.对其18SrRNA基因进行了克隆测序并登录GenBank(AY758384).依据18SrRNA基因建立的系统进化树表明:该菌与Schizochytriumlimacinum具有紧密的亲源关系. 图7表2参29  相似文献   

18.
Dover sole (Solea solea, Linneaus 1758) were raised from first feeding on brine shrimp (Artemia sp.) with different contents and compositions of the essential fatty acids (EFA) arachidonic acid (ARA, 20:4n − 6); eicosapentaenoic acid (EPA, 20:5n − 3), and docosahexaenoic acid (DHA, 22:6− 3), and their metabolic rate and tolerance to hypoxia measured prior to and following metamorphosis and settlement. Four dietary Artemia preparations were compared: (1) un-enriched; (2) enriched with a commercial EFA mixture (Easy DHA SELCO Emulsion); (3) enriched with a marine fish oil combination (VEVODAR and Incromega DHA) to provide a high ratio of ARA to DHA, and (4) enriched with these fish oils to provide a low ratio of ARA to DHA. Sole fed un-enriched Artemia were significantly less tolerant to hypoxia than the other dietary groups. Larvae from this group had significantly higher routine metabolic rate (RMR) in normoxia, and significantly higher O2 partial pressure (PO2) thresholds in progressive hypoxia for their regulation of RMR (P crit) and for the onset of agitation, respiratory distress and loss of equilibrium. Metamorphosis was associated with an overall decline in RMR and increase in P crit, but juveniles fed on un-enriched Artemia still exhibited higher P crit and agitation thresholds than the other groups. Sole fed un-enriched Artemia had significantly lower contents of EFA in their tissues, both before and after settlement. Thus, enriching live feeds with EFA has significant effects on the respiratory physiology of sole early life stages and improves their in vivo tolerance to hypoxia. We found no evidence, however, for any effect of the ratio of ARA to DHA.  相似文献   

19.
柴油污染土壤生物修复对土壤酶活性的影响   总被引:2,自引:0,他引:2  
在柴油污染土壤的生物修复过程中,分析了土壤中柴油降解菌数量和3种土壤酶活性(过氧化氢酶、脱氢酶和脂酶)等生物活性指标与土壤中柴油去除率的相关性.结果表明,土壤受到柴油污染后的一段时间后过氧化氢酶、脱氢酶和脂酶的活性上升,而后随着土壤中石油烃的降解,脂酶又不断降低.进一步分析表明脂酶活性与柴油降解率及柴油降解菌数量都具有很好的正相关性,可以采用土壤脂酶活性来指示柴油生物降解成效.  相似文献   

20.
Larval fish growth and survival depends not only on prey quantity, but also on prey quality. To investigate effects of prey fatty acid concentration on larval herring growth, we collected different prey organisms and larval herring (Clupea harengus L.) in the Kiel Canal during the spring season of 2009. Along with biotic background data, we analysed fatty acids both in prey organisms and in the larvae and used biochemically derived growth rates of the larvae as the response variable. Larval herring reached their highest RNA/DNA derived growth rates only at high docosahexaenoic acid (DHA) concentration. When the ratio of copepodids to lesser quality cirriped nauplii was low, larval growth and larval DHA concentration were both significantly negatively affected. This was true even as prey abundance was increasing. This finding indicates that even in mixed, natural feeding conditions, growth variations are associated with DHA availability in larval fish.  相似文献   

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