前体物质对辅酶Q_(10)生物合成的影响

首次研究了以辅酶Q10 (CoQ10 )主要侧链供给前体物质———茄尼醇和醌环供给前体———羟基苯甲酸和辅酶Q0 对粟酒裂殖酵母(Schizosaccharomycespromb 2 1794 - 2 3) 生产CoQ10 产量的影响,并对前体的转化工艺进行了初步研究。确定了适宜于粟酒裂殖酵母生长及高转化前体生成CoQ10 的条件为:酵母在2 8℃下,2 2 0r/min于发酵培养基中培养18h后,加入0 5 g/L茄尼醇继续发酵培养18h ,进行前体转化反应。结果表明,单独添加茄尼醇能达到最大产量33 1mg/L ,比对照样品增加了91% ,任2种前体物质共同添加都要比单独添加茄尼醇时产量低。茄尼醇和CoQ0 共同添加时,单位细胞胞外辅酶CoQ10 的产量达到最高的1 35mg/g ,比对照样品增加了117%。     

酵母转化茄尼醇生成辅酶Q10的超临界CO2体系条件

超临界CO2作为非水相介质对酵母转化茄尼醇生成辅酶Q10的底物和产物具有良好的溶解作用,但对微生物的存在会产生一定影响,选择合适的超临界CO2体系条件有利于辅酶Q10产量的提高。通过研究超临界CO2流体的压强、酵母细胞在超临界CO2流体中的反应时间、添加介质种类以及添加量等条件对粟酒裂殖酵母细胞活性和辅酶Q10产量的影响,确定对酵母细胞生存影响较小并显著促进辅酶Q10生产的超临界CO2体系条件为:超临界CO2压强为10.5 MPa,反应时间为16 h,向菌体中添加菌体湿重3倍的培养基,超临界CO2流体中辅酶Q10的产量达到55.71μg/mL。比液体深层发酵培养辅酶Q10产量提高97.5%。研究结果表明,通过优化酵母转化茄尼醇生成辅酶Q10的超临界CO2体系条件,可以显著提高酵母细胞生成辅酶Q10的能力。     

生物降酸生产笃斯越桔全汁酒的研究

以笃斯越桔为原料,采用粟酒裂殖酵母降酸生产全汁越桔酒,通过正交实验确定酒精发酵的最佳条件为:发酵温度28℃.发酵时间为5d,酵母添加量0.5%.生物降酸的最佳发酵条件为:粟酒裂殖酵母接种量5%,发酵温度25℃,发酵时间4d.澄清剂的最佳使用量为:明胶200me/L和皂土2000mg/L澄清效果最佳.     

酿酒酵母静息细胞转化合成2-苯乙醇

利用酿酒酵母静息细胞转化合成2-苯乙醇,确定了静息细胞转化菌龄、转化液缓冲液体系和辅酶NADH再生辅助底物。在单因子试验基础上,用Plackett-Burm设计从影响转化6个因素中筛选出影响细胞转化的显著因素:辅助底物乙醇质量浓度、转化液pH和转化温度,以BoxBenhnken试验设计结合响应面法分析确定显著因子的最优水平。结果表明,最佳转化条件为乙醇质量浓度16.5 g/L、pH 5.1、转化温度26℃,2-苯乙醇产量为3.49 g/L(转化率58.8%),比优化前提高了49.14%,静息细胞重复使用8次,2-苯乙醇产量无明显下降。转化液中加入10%的大孔吸附树脂对产物进行原位产物吸附,转化40 h,2-苯乙醇总质量浓度达9.34 g/L,L-苯丙氨酸转化率为83.9%,产量比未加入大孔树脂提高了171.5%。     

唾液链球菌嗜热亚种Y-2细胞转化法制备γ-氨基丁酸

研究反应pH值、反应温度、重金属盐、表面活性剂、底物浓度、菌体质量浓度和磷酸吡哆醛添加量对Streptococcus salivarius subsp.thermophilus Y-2细胞转化法生产γ-氨基丁酸的影响。获得反应体系的最佳组成为:湿菌体25g/L、BaCl2 40mmol/L、Triton X-100体积分数0.02%、L-谷氨酸单钠盐(L-monosodium glutamate,MSG)47.5g/L和L-谷氨酸(L-glutamic acid,L-Glu)90.0g/L。该体系在40℃、pH 4.5和搅拌速度100r/min的最适转化条件下进行反应72h,转化液GABA产量达到了(87.16±4.33)g/L,细胞平均生产力为(48.42±2.41)mg/(h.g),摩尔转化率为(97.60±4.71)%。     

重组谷氨酸脱羧酶大肠杆菌合成γ-氨基丁酸条件的优化

研究重组谷氨酸脱羧酶大肠杆菌合成γ-氨基丁酸(γ-aminobutyric acid,GABA)的适宜条件。检测温度、p H值、表面活性剂、金属离子、底物与菌体质量比以及反应体系体积对GABA转化效率的影响。结果表明:最优转化条件为:转化体系5 m L、底物L-谷氨酸钠浓度0.1 mol/L、重组大肠杆菌细胞6.4 mg(干质量)、Triton-100体积分数0.06%、Ca2+浓度0.6 mmol/L,转化温度45℃、反应体系p H 4.5。在该体系下反应7 h,GABA合成量达到26.1 g/L,GABA转化效率在1 h时达到最高,为13.8 g/(g h),较优化前提高1.5 倍。     

黑曲霉B1401生物法转化单宁酸制备没食子酸的研究

实验优化了黑曲霉B1401液态发酵直接投加单宁酸生物法制备没食子酸的工艺条件,得出的最佳工艺条件为：底物单宁酸添加方式为分批添加,其累计添加质量浓度达204 mg/mL;转化过程pH维持在4.0;转化温度为30 ℃;转化过程摇瓶转速为160 r/min。将优化后的工艺条件放大到5 L发酵罐,单宁酸转化效率优于摇瓶发酵,转化平衡时间由摇瓶的48 h缩短至44 h,没食子酸的产率由70.35%提高至74.04%,质量浓度由144 mg/mL提高至161 mg/mL。     

重组大肠杆菌双相体系合成(R)-2-羟基-3-苯基丙酸

在水/正己烷双相体系中,利用重组大肠杆菌全细胞催化苯丙酮酸(PPA)提高(R)-2-羟基-3-苯基丙酸(PLA)的产量。研究了双相体系中正己烷的体积分数、转化温度、转速、底物浓度和菌体量对PLA产量的影响。结果表明:最优转化条件为正己烷体积分数为40%;转化温度为40℃;转化转速为300 r/min;底物浓度为15 g/L;湿菌体浓度为15 g/L。在此条件下进行全细胞转化,PLA的产率为(7.25±0.08)g/(L·h)。分批补加底物,生成PLA的单位时间产量为(8.77±0.13)g/(L·h)。     

10-羟基-12-十八碳烯酸的静息细胞合成

10-羟基-12-十八碳烯酸(10-HOE)是一种典型的微生物源羟基不饱和脂肪酸,具有抑菌、抗炎、抗过敏等功效,在食品、药品、化工等行业具有很好的应用前景,因此,10-HOE的生物合成十分重要。本文以亚油酸为底物,以含10-亚油酸水合酶的重组大肠杆菌为催化剂,采用静息细胞法生物合成10-HOE,通过优化黄素腺嘌呤二核苷酸(FAD)浓度、溶剂种类及含量、甘油含量、氯化钠浓度、转化温度、溶液pH值、菌浓度、底物浓度和培养时间等9个单因素转化实验,来提高10-HOE的转化率。细胞转化生物合成的优化条件为:FAD浓度0.25 mmol/L,添加2%的正己烷,氯化钠浓度15 mmol/L,亚油酸底物质量浓度20 g/L,菌质量浓度200 g/L、溶液pH7.0,在37 ℃、200 r/min的摇床中培养24 h,获得的10-HOE转化率为(73.1±0.9)%,转化质量浓度为(16.3±0.2) g/L。该转化是目前为止10-HOE生物合成报道的最高转化浓度。本研究为大规模制备10-HOE提供了工艺路线,为工业化生产10-HOE提供参考。     

优选适于发酵玫瑰香葡萄酒的粟酒裂殖酵母

为筛选出发酵玫瑰香葡萄酒效果最佳的粟酒裂殖酵母菌株,该试验首先通过不同发酵温度（23、25、28℃）和不同接菌量（1×106、5×106、1×107CFU/mL）对玫瑰香葡萄进行发酵,优选出适用于粟酒裂殖酵母的最适发酵温度和最适接菌量。然后利用前期根据发酵动力学试验筛选出的16株粟酒裂殖酵母对玫瑰香葡萄进行单独发酵。结果表明：适合粟酒裂殖酵母的发酵温度为25℃,接菌量为5×106CFU/mL。相较于其他发酵菌株,筛选出的粟酒裂殖酵母Sp-410单独发酵玫瑰香葡萄酒的苹果酸的降解率（97.98%）和色度值（1.26）最高,乙酸含量（0.18 g/L）和乙醛含量（48.46 mg/L）最低,因此,优选出的粟酒裂殖酵母菌Sp-410为酿造优质玫瑰香葡萄酒的优良菌种。     

吐温80增溶-紫外分光光度法测定辅酶Q_(10)脂质体的载量及包封率

在辅酶Q10脂质体的制备过程中,载量和包封率是评价辅酶Q10脂质体的2个重要质量指标。采用表面活性剂吐温80对辅酶Q10脂质体进行增溶,再结合紫外分光光度法测定其载量和包封率。研究结果表明,辅酶Q10浓度在2.5～50μg/mL范围内,吐温80增溶法与以乙醇为溶剂的反相高效液相色谱法以及紫外分光光度法有良好的相关性(R2>0.999);空白脂质体中,辅酶Q10的加样回收率在(98.26±0.63)%～(101.20±1.28)%之间,相对标准偏差RSD<2%(n=6);该法用于测定辅酶Q10脂质体中总辅酶Q10含量的RSD<5%(n=6);不同载量[(3.22±0.01)%～(13.62±0.31)%]的辅酶Q10脂质体的包封率均高于95%(RSD<1%,n=6)。与以乙醇为溶剂的反相高效液相色谱法以及紫外分光光度法相比,该法具有准确可靠、简单、重现性较好的优点。     

高效液相色谱法测定蜂王幼虫中辅酶Q10含量

建立一种用高效液相色谱测定蜂王幼虫中辅酶Q10 含量的方法。蜂王幼虫样品研磨均质后,经无水乙醇-正己烷液/ 液萃取,提取液经浓缩后,无水乙醇定容。以甲醇为流动相,采用XBridge shield RP18 柱分离,在紫外检测器275nm 波长处对样品中的辅酶Q10 进行测定。辅酶Q10 在12min 内得到较好的分离,方法线性良好,回收率为95.0%～101.8%,相对标准偏差(RSD)小于8%,检出限为0.23mg/L。该方法操作简单、准确、可靠、灵敏度高,适用于测定蜂王幼虫中辅酶Q10 的含量,已被用于分析一些实际样品,调查辅酶Q10 在蜂王幼虫样品中的含量情况。     

Analysis of coenzyme Q10 in bee pollen using online cleanup by accelerated solvent extraction and high performance liquid chromatography

A method for the determination of coenzyme Q10 in bee pollen has been developed applying an online cleanup of accelerated solvent extraction and using environmentally acceptable organic solvents. The extracted samples were analysed by high performance liquid chromatography with diode array detection. The optimised method employed 10 mL extraction cells, 1 g sample size, absolute ethanol as extraction solvent, 80 °C of extraction temperature, one extraction cycle, 5 min of static time, Cleanert Alumina-N as sorbent and 60% flush volume. The method was validated by means of an evaluation of the matrix effects, linearity, limit of detection (LOD) and quantification (LOQ), trueness, precision and stability. The assay was linear over the concentration range of 0.25–200 mg/L and the LOD and LOQ were 0.16 and 0.35 mg/kg, respectively. The recoveries were above 90%. The inter- and intra-day precision was below 6.3%. The method has been successfully applied to the analysis of bee pollen samples. For 20 bee pollen products, the coenzyme Q10 content varied from not detectable to 192.8 mg/kg.     

响应曲面法优化辅酶Q10产生菌发酵培养基

以辅酶Q10产生菌R-SL15为实验菌株,为提高其辅酶Q10产量,对其进行培养基优化,得到最优发酵培养基。采用Plackett-Burman实验设计和Box-Behnken响应面分析方法对R-SL15的培养基进行优化模型的建立,得出最优发酵培养基为:葡萄糖18.90g/L,酵母粉5.54g/L,(NH4)2SO40.98g/L,KH2PO40.95g/L,牛肉膏6g/L,MgSO4.7H2O0.75g/L,FeSO4.7H2O100mg/L,NaCl10g/L,蒸馏水1L。辅酶Q10产量为51.31mg/L,比优化前的25.74mg/L提高了99.34%。该回归模型高度显著(R2=0.9423),拟合性好,可用于预测。     

阿魏酸类似物对强的松诱导的小鼠氧化应激的影响

本实验研究阿魏酸及其类似物姜黄素、谷维素对小鼠体内氧化应激的影响。将90 只雄性ICR小鼠随机分为6 组：正常组、模型组、阿魏酸组、姜黄素组、谷维素组和阳性对照（辅酶Q10）组。每3 d正常组肌肉注射生理盐水,其他组注射40 mg/kg mb强的松进行造模。正常组和模型组饲喂正常饲料,其他4 组每千克饲料中分别添加40 mg的阿魏酸、姜黄素、谷维素和辅酶Q10。饲养4 周后开始进行行为学实验（糖水偏好和强迫游泳实验）,饲养8 周后处死小鼠,并提取心、肝、腿部肌肉的线粒体,测定线粒体、血浆、肝脏、心脏和肌肉组织中的氧化应激相关指标。结果表明：1）阿魏酸组小鼠体质量较初始体质量明显下降;辅酶Q10组小鼠的糖水偏好程度显著高于模型组和阿魏酸组（P＜0.05）。姜黄素、谷维素和辅酶Q10组小鼠在强迫游泳实验中的绝望状态时间均显著短于模型组（P＜0.05）。2）阿魏酸、姜黄素、谷维素和辅酶Q10小鼠下丘脑促肾上腺素释放激素和垂体促肾上腺皮质激素的含量均显著低于模型组（P＜0.05）。谷维素和辅酶Q10组小鼠血浆皮质酮的含量显著低于模型组（P＜0.05）。3）谷维素组和辅酶Q10组血浆和组织中的丙二醛含量显著低于模型组（P＜0.05）,过氧化氢酶活力显著高于模型组（P＜0.05）。4）谷维素和辅酶Q10组的组织中总超氧化物歧化酶活力和肌肉线粒体中Mn-SOD活力显著高于模型组（P＜0.05）。5）谷维素组和辅酶Q10组线粒体膜电位显著高于模型组（P＜0.05）,辅酶Q10组线粒体中NADH/NAD＋比值和乙酰辅酶A含量显著高于模型组（P＜0.05）。在本实验条件下,综合抗应激和组织线粒体抗氧化能力由小到大依次为阿魏酸＜姜黄素＜谷维素＜辅酶Q10。对于提高组织中抗氧化酶的活力和改善氧化应激造成的线粒体功能紊乱,谷维素和辅酶Q10的效果显著大于阿魏酸和姜黄素,其中阿魏酸抗氧化能力最差,辅酶Q10抗氧化能力最高,谷维素抗氧化能力接近于辅酶Q10。经过初步判断,阿魏酸及其类似物的抗氧化能力与其亲脂性相关。     

In vitro bioaccessibility of coenzyme Q10 in enriched yoghurts

In this study, the aim was to determine the bioaccessibilities of coenzyme Q10 (CoQ10) in yoghurt samples produced using enriched skim milk with different coenzyme Q10 preparations. First, emulsified coenzyme Q10, γ‐cyclodextrin/coenzyme Q10 complex and nanoparticle coenzyme Q10 were prepared with the reference coenzyme Q10 standard. These coenzyme Q10 preparations were added into the milk to produce coenzyme Q10 enriched yoghurt samples. Nanoparticle coenzyme Q10 was obtained as in spherical shape with 176.00 ± 50.62 nm diameter. Coenzyme Q10 bioaccessibility was found as 50.59 ± 1.88% in control yoghurt. The yoghurt enriched nanoparticle coenzyme Q10 had the highest coenzyme Q10 bioaccessibility (73.81 ± 1.61%) among the produced yoghurts (P < 0.01). Coenzyme Q10 bioaccessibilities were also found as 63.75 ± 0.91% and 46.83 ± 1.27% in yoghurts enriched with emulsified coenzyme Q10, and γ‐cyclodextrin/coenzyme Q10 complex, respectively.     

Control of melanosome transfer by promoting shrinkage or expansion of melanocyte dendrites

Coenzyme Q₁₀ is an endogenous antioxidant found in the skin along with vitamins A, C and E. Coenzyme Q₁₀ is used increasingly in cosmetic products and is advertised as a skin energizer, protector against skin aging, skin repairer and an anti-wrinkling agent. As the outermost layer of skin, the stratum corneum is the interface between the body and the environment and requires antioxidants to protect it and the epidermis and dermis below it. Nutrient levels in the stratum corneum correlate with those in the skin. Two forms of coenzyme Q₁₀ (pure coenzyme Q₁₀ and a yeast-based coenzyme Q₁₀) were investigated in a commercial vehicle. Two groups of subjects were tested aged 21–29 and 51–70, respectively. Coenzyme Q₁₀ absorption in the stratum corneum was determined after a 1-h application and ethanol extraction. Significantly more yeast coenzyme Q₁₀ was absorbed than the pure coenzyme Q₁₀ and the middle-aged subjects absorbed about twice as much coenzyme Q₁₀ as did the younger subjects. Skin antioxidants were significantly increased by yeast coenzyme Q₁₀ but not by pure coenzyme Q₁₀. Peroxides declined in the stratum corneum after twice daily application of coenzyme Q₁₀ with both forms but the decrease was greater with the yeast form. The older subjects had significantly higher baseline levels of lipid peroxides than did the younger group, indicating an increase in skin oxidative damage with age. The yeast coenzyme Q₁₀ is the superior form of coenzyme Q₁₀ for human skin application.     

Effects of coenzyme Q10, α-tocopherol and ascorbic acid on oxidation of cholesterol in chicken liver pâté

The aim of this study was to determine whether supplemental addition of coenzyme Q₁₀ and ascorbic acid or α-tocopherol, either alone or together, can prevent oxidative damage in chicken liver pâté, as reflected by reduced formation of cholesterol oxidation products (COPs) and by preservation of sensorial quality. Separate groups of chicken liver pâtés had no supplements (control) or were supplemented with coenzyme Q₁₀ (0.2 g/kg) and either ascorbic acid (2 g/kg) or α-tocopherol (0.2 g/kg), or both. All products were pasteurised (82 °C) or sterilised (121 °C). Four COPs were found: 7α-, 7β-, 20α- and 25-hydroxycholesterol. The COP radical scavenger function of coenzyme Q₁₀ (control, 5.16 mg/kg; plus Q₁₀, 3.94 mg/kg) and the synchronous actions of coenzyme Q₁₀ and α-tocopherol (2.6 mg/kg) were confirmed in sterilised pâtés. Generally, in pasteurised and sterilised pâtés, the most efficient scavenger function was with ascorbic acid either alone or together with α-tocopherol, where the formation of COPs was below the limit of detection. An increase of 1.9 mg/kg in COP production during heating was also seen in samples without added antioxidants. There was a weak interdependence between the content of COPs and the sensory parameters of the pâté. For addition of antioxidants, in the pasteurised pâté, colour and smell were slightly improved, but flavour deteriorated; in the sterilised pâté, colour was slightly worse, with a more tender texture. Overall, instrumentally measured colour and sensory properties (except texture) showed no significant differences between pasteurisation and sterilisation.     

辅酶Q_（10）软胶囊中的辅酶Q_（10）与V_E高效液相色谱法研究

本研究建立了含天然VE的辅酶Q10软胶囊中的辅酶Q10与天然维生素E同时测定的方法。该方法采用高效液相色谱法,以Agilent TC-C18（2）色谱柱（250mm×4.6mm,5μm）为分析柱;以甲醇：乙醇（75：25）为流动相;检测波长为290nm。结果表明,本方法测定辅酶Q10的平均回收率为：98.40%,RSD为：0.86%（n=6）;天然VE的平均回收率为：101.69%,RSD为：0.58%（n=6）。该方法简单、方便、准确、可靠、可用于同时测定同一制剂中共存的辅酶Q10与VE。