新型膳食补充剂吡咯喹啉醌生物合成及菌株选育研究进展

吡咯喹啉醌（Pyrroloquinoline Quinone,PQQ）是继烟酰胺核苷酸（NAD、NADP）和核黄素核苷酸（FMN、FAD）后新发现的一种具有高度水溶性和稳定性的芳香族三环邻醌化合物，它广泛存在于微生物、植物、动物和人体中。作为一种类维生素辅酶，吡咯喹啉醌具有多种生理功能，在促进机体生长、防护肝损伤、促进神经生长因子合成、调节机体自由基水平等方面发挥巨大作用。由于PQQ在营养健康方面的积极效果，吡咯喹啉醌二钠盐已被美国和欧盟作为新型的膳食补充剂广泛推广，其在医药、食品、农业等方面具有广阔的应用前景，因此未来应深入研究如何提高吡咯喹啉醌高产菌株产量。本文对国内外PQQ高产菌株的选育及生物合成代谢途径的研究进行了总结，对PQQ生物合成的基因表达及菌株改造进行了展望。     

生物法制备乙醇酸的研究

目的:对氧化葡萄糖酸杆菌生物转化乙二醇生产乙醇酸的工艺进行研究.方法:首先采用单因素试验优化了氧化葡萄糖酸杆菌生物转化乙二醇生成乙醇酸的工艺,在此基础上通过正交试验对转化工艺进行了进一步探讨.结果:氧化葡萄糖酸杆菌生物转化乙二醇生成乙醇酸的最优工艺为12.6%静息细胞、5.6%乙二醇维持pH5.84转化48h,乙二醇单体转化率可达60.886%.结论:应用氧化葡萄糖酸杆菌生物转化乙二醇生产乙醇酸的工艺的切实可行.     

生物电化学法转化甘油生产1,3-二羟基丙酮

氧化葡萄糖酸杆菌中依赖辅基吡咯喹啉醌(Pyrroloquinoline Quinone,PQQ)的膜结合甘油脱氢酶(Glycerol dehydrogenase,GDH)是酶法转化甘油生成1,3-二羟基丙酮(1,3-Dihydroxyacetone,DHA)的关键酶。以铁氰化钾为电子媒介体,采用生物电化学法,再生甘油脱氢酶的辅基PQQ,从而实现酶法循环转化甘油生产DHA。设计电耦联反应装置,在(28±2)℃,370mV电压下反应18h,DHA质量浓度达到27.21g/L,甘油转化率为52.93%。     

VC混菌发酵中大菌不同胞外组分对小菌的影响

在维生素C(VC)二步混菌发酵中,巨大芽孢杆菌可显著促进氧化葡萄糖酸杆菌产VC前体2-酮基-L-古龙酸(2-KLG)。采用硫酸铵分级盐析沉淀、柱层析及电泳技术对巨大芽孢杆菌胞外液中的活性物质进行了分离纯化。通过测定氧化葡萄糖酸杆菌生长细胞转化L-山梨糖为2-KLG的生成量、产酸菌关键酶L-山梨糖脱氢酶(SDH)酶活性及产酸菌的活菌数,研究了VC两步发酵中大菌不同胞外组分对小菌的作用。结果表明:大菌胞外36 000和44 300两个组分蛋白质对小菌产酸、SDH酶活具有促进作用。     

利用柿子皮混菌发酵制备果醋及品质分析

以柿子加工副产物柿子皮为主要原料,采用氧化葡萄糖酸杆菌、植物乳杆菌、明串珠菌发酵酿造果醋。通过单一菌种、多菌组合的发酵实验,以发酵时间和产品质量为指标,确定混合菌种发酵的最佳组合。采用顶空固相微萃取结合气相色谱-质谱联用技术鉴定了最佳醋品的挥发性香气成分,采用反相高效液相色谱法测定了主要有机酸含量。结果表明：当V（氧化葡萄糖酸杆菌）∶V（植物乳杆菌）＝1∶1时,醋品品质最佳,发酵时间6 d,感官品评94.90 分,共鉴定出51 种挥发性香气成分,主要为醇类和酯类,有机酸中乳酸和乙酸为主要有机酸,柿香味和醋香浓郁、协调,口感佳,品质优。     

微生物发酵法生产D-木糖酸

D-木糖酸是一种多功能的平台化合物。它可以作为高效水泥黏合剂,也可以作为合成多种化合物的前体,比如共聚酰胺类、多酯类、水凝胶类和1,2,4-丁三醇。随着葡萄糖的价格越来越高,D-木糖酸或许可以代替葡萄糖酸。目前,大规模的生产还没有发展起来,这反映了对于D-木糖酸的市场限制。以D-木糖为底物利用氧化葡萄糖酸杆菌发酵生产D-木糖酸,可以得到高得率,这使得氧化葡萄糖酸杆菌成为一种生物技术生产D-木糖酸的不错选择。氧化葡萄糖酸杆菌可以直接利用植物废料水解液生产D-木糖酸,但是生产速率和产量都不高。这是因为氧化葡萄糖酸杆菌对水解液抑制物敏感。近来,D-木糖酸已经可以通过基因工程改造后的大肠杆菌、酿酒酵母和克鲁维酵母生产。随着进一步的发展,基因工程菌可能会为大规模生产D-木糖酸提供新的选择。     

氧化葡萄糖酸杆菌培养及转化1,2-丙二醇生产D-乳酸研究

对氧化葡萄糖酸杆菌培养及转化1,2-丙二醇生产D-乳酸进行研究.采用单因素实验优化了氧化葡萄糖酸杆菌的生长培养基,然后通过不同转化条件对该菌转化1,2.丙二醇生成D-乳酸的工艺进行了初步探讨.氧化葡萄糖酸杆菌生长的最佳碳源为6%山梨醇、最佳氮源为3%酵母粉;转化1,2-丙二醇的最优工艺为:10%静息细胞、20%(R,S)1,2-丙二醇维持pH6.0,250mL摇瓶装液30mL,28℃、220r/min培养30h.     

3 种脂环酸芽孢杆菌分离培养基的性能评价

目的：比较3 个厂家的K培养基、酵母粉淀粉葡萄糖（yeast extract starch glucose medium,YSG）培养基和耐酸耐热芽孢菌（Alicyclobacillus）培养基（Bacillus acidoterrestris medium,BAT）对脂环酸芽孢杆菌的检测效果。方法：选用标准菌株、分离株及实际样品对3 个厂家（广东环凯微生物科技有限公司（简称HK,下同）、厂家Ⅰ和厂家Ⅱ）的K培养基、YSG培养基和BAT培养基进行生长菌落数及分离效果的评估。结果：酸土脂环酸芽孢杆菌标准株和脂环酸芽孢杆菌分离株4-2在K、YSG和BAT 3 种培养基上生长的菌落数（3 个厂家培养基上的菌落数平均数）均无显著性差异（P＞0.05）;酸热脂环酸芽孢杆菌标准株和脂环酸芽孢杆菌分离株5-3在K培养基上不生长,在YSG培养基和BAT培养基上生长良好,其菌落平均数统计学上无显著性差异（P＞0.05）;对实际样品中酸土脂环酸芽孢杆菌的分离,3 个厂家的同种培养基分离率和符合率相当;对实际样品中酸热脂环酸芽孢杆菌的分离,在YSG和BAT培养基上,HK和厂家Ⅱ的分离率和符合率相当,优于厂家Ⅰ。结论：K培养基不适合所有脂环芽孢杆菌,但分离酸土脂环酸芽孢杆菌效果很好,YSG培养基和BAT培养基适合所有脂环酸芽孢杆菌。HK和厂家Ⅱ的培养基优于厂家Ⅰ。     

黑曲霉AnM1产葡萄糖酸及纯化低聚异麦芽糖

该文对黑曲霉AnM1摇瓶发酵产葡萄糖酸及利用黑曲霉AnM1纯化低聚异麦芽糖进行研究。根据葡萄糖酸生产强度和得率确定黑曲霉AnM1摇瓶发酵培养基。发酵后收集的黑曲霉AnM1菌体可重复利用产葡萄糖酸,重复利用3次后葡萄糖酸的生产强度仍在7 g/（L·h）以上,得率为1.02 g/g葡萄糖。利用黑曲霉AnM1菌体氧化低聚异麦芽糖反应液中的葡萄糖,可将低聚异麦芽糖含量提高至总糖量的90%以上,其中有效三糖（异麦芽糖、潘糖、异麦芽三糖）含量占总糖的60%以上。黑曲霉AnM1具有较好的葡萄糖酸生产能力,菌体可用于纯化低聚异麦芽糖并能重复使用,在葡萄糖酸和低聚异麦芽糖生产方面具有很好的应用前景。     

吡咯喹啉醌研究新进展

吡咯喹啉醌（pyrroloquinoline quinone,PQQ）是细菌脱氢酶中的一种氧化还原辅助因子,又是一种抗氧化剂,能避免细胞内氧化反应以及体外生物活性物质产生活性氧导致的细胞损伤,为细胞的生长发育提供营养与维生素,同时使细胞具有抗氧化的耐受性。它对植物病原真菌起到生物控制剂的作用,能诱导蛋白激酶参与哺乳动物细胞分化发育过程。PQQ能通过增加不溶性磷酸盐的利用率来提高作物产量,它与氧化还原循环功能有很强相关性,具有抗神经退行性、抗癌、信号传导等功能。     

The thermostability, bioactive compounds and antioxidant activity of some vegetables subjected to different durations of boiling: Investigation in vitro

This research was performed in order to compare the water and acetone extracts of raw and boiled for 10, 20, 40 and 60 min Korean lotus roots (KLR) and Polish white onion (PWO) in the contents of their bioactive compounds, antioxidant activity and thermostability.It was found that polyphenols (mg GAE/g), flavanols (μg GAE/g), flavonoids (mg CE/g), anthocyanins (mg CGE/kg) and tannins (mg CE/g) in water extract of raw lotus roots were 14.18 ± 0.7, 8.41 ± 0.5, 1.09 ± 0.06, 21.3 ± 1.2 and 7.29 ± 0.4, and of white onion - 11.11 ± 0.6, 6.78 ± 0.3, 0.71 ± 0.03, 17.00 ± 0.9 and 1.64 ± 0.08, respectively, and significantly higher in KLR (P < 0.05). The antioxidant activity of raw KLR water extract (139.4 ± 6.1, 53.1 ± 3.6 and 89.3 ± 4.6 μmol TE/g for DPPH, CUPRAC and ABTS, respectively) was significantly higher than in white onion (23.84 ± 1.8, 31.9 ± 2.1 and 38.14 ± 2.6 for DPPH, CUPRAC and ABTS, respectively, P < 0.05).The thermostability of the water KLR extract’s of polyphenols, flavanols, flavonoids, anthocyanins and tannins was high and even after 60 min of boiling remains as 40.0, 42.3, 50.5, 41.4 and 41.0%, respectively. After 60 min of boiling the most thermostable compounds were flavonoids - remaining at 50.5% in water extract of KLR. Also after 60 min of boiling the thermostability of the antioxidant activity of water extracts of KLR remained significantly high: 40.6, 42.3, 46.3 and 43.6%, according to DPPH, FRAP, ABTS and CUPRAC assays, respectively.Similar relationship was obtained with acetone extracts, but the value was lower than with the water ones. In conclusion, the contents of some bioactive compounds, the antioxidant activity and the thermostability in water and acetone extracts of KLR are significantly higher than the same indices in PWO. FTIR and fluorimetry can be used as additional markers for the characterization of bioactive compounds in vegetables.     

Nutritive value of masau (Ziziphus mauritiana) fruits from Zambezi Valley in Zimbabwe

Ziziphus mauritiana (masau) fruits are consumed by many people in Zimbabwe. The fruits contribute significantly to people’s diet when they are in season. The objective of this study was to determine the nutritional content of the fruits and, hence, quantify their contribution to the diet. Samples of masau were collected in two seasons (August 2006 and August 2007). Both macronutrients and micronutrients were determined using standard AOAC methods of analysis. Dry matter content ranged from 21.1 ± 0.2 to 24.1 ± 0.3 g 100 g⁻¹ of edible portion of the sweet and sour fruits, and 84.8 ± 0.2 to 87.2 ± 0.2 g 100 g⁻¹ for the dried fruit. Crude protein per 100 g edible portion of dry weight ranged between 7.9 ± 0.0 and 8.7 ± 0.0 g, crude fat from 0.8 ± 0.0 to 1.5 ± 0.0 g, crude fibre from 4.9 ± 0.0 to 7.3 ± 0.0 g, ash between 3.0 ± 0.0 and 4.3 ± 0.0 g and carbohydrate between 79.5 ± 0.0 and 83.2 ± 0.0 g. The fruits were rich in vitamin C (15.0 ± 0.0–43.8 ± 0.02 mg 100 g⁻¹) and the energy values ranged between 1516.0 ± 1.73 and 1575.0 ± 2.3 kJ 100 g⁻¹. Furthermore, the fruits contained (mg 100 g⁻¹ of dry weight) potassium from 1865.0 ± 1.3 to 2441.0 ± 1.1, calcium from 160.0 ± 0.3 to 254.0 ± 0.1, sodium between 185.0 ± 0.1 and 223.0 ± 0.2, magnesium between 83.0 ± 0.0 and 150.0 ± 0.13 and phosphorous from 87.0 ± 0.1 to 148.0 ± 0.5. Manganese and copper contents ranged between 0.7 ± 0.03 and 1.6 ± 0.03, while iron and zinc ranged between 2.1 ± 0.43 and 4.3 ± 0.1, and 0.6 ± 0.0–0.9 ± 0.0 mg 100 g⁻¹ of dry weight, respectively. The masau fruit is therefore a good potential source of carbohydrates, proteins and micronutrients, such as calcium, potassium, sodium, phosphorous, copper, iron, Vitamin C and zinc.     

Evaluation of culture media for enumeration of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium animalis in the presence of Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus

The study compared the growth capability of probiotic (Lactobacillus acidophilus La05, Lactobacillus casei Lc01 and Bifidobacterium animalis Bb12) and non-probiotic (Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus) cultures on twenty-one culture media grouped according to selectivity: non-selective agars, selective agars without antibiotics and MRS agars containing different combinations of lithium chloride, cystein, bile salts and antibiotics. Four of these media were selected for quantitative enumeration of L. acidophilus La05, L. casei Lc01, and B. animalis Bb12. The best culture media and incubation conditions for enumeration of the probiotic cultures were: B. animalis: MRS agar with dicloxacillin, 37 °C or 42 °C, anaerobiosis; L. acidophilus: MRS agar with bile salts, 37 °C or 42 °C, aerobiosis; L. casei: MRS agar with lithium chloride and sodium propionate, 37 °C or 42 °C, aerobiosis or anaerobiosis. Plating on MRS with glucose replaced by maltose, 37 °C or 42 °C, anaerobiosis, will distinguish probiotic from non-probiotic cultures. For enumeration of each probiotic in a mixed culture, the following media and incubation conditions were recommended: B. animalis: 4ABC-MRS, 42 °C, anaerobiosis, L. acidophilus: LC medium, 42 °C, aerobiosis or anaerobiosis and L. casei: LP-MRS, 42 °C, aerobiosis or anaerobiosis. In all experiments, differences in counts using pour plating or surface plating were not significant (P ≤ 0.05).     

Quality assessment of wild European eel (Anguilla anguilla) stored in ice

The quality assessment of wild European eel (Anguilla anguilla) stored in ice and in boxes without ice (3 ± 1 °C) was investigated by the sensory analysis, levels of nucleotide breakdown products and biogenic amines for up to 19 days. Sensory analysis was assessed using the Tasmanian Food Research Unit Scheme. K and related values (K_i, G, P, H and F_r) were used as freshness indicators. Linear regressions (r²) obtained from K, K_i, G, P, H and F_r were 0.95, 0.96, 0.83, 0.96, 0.99 and 0.96, respectively, for eel stored in ice whereas, for eel kept in boxes without ice, the values were 0.86, 0.86, 0.96, 0.91, 0.98 and 0.86, respectively. When eel stored in ice and in boxes without ice were considered at the limit of acceptability by assessors at ∼12–14 days and ∼5–7 days, respectively, the average K, K_i and P values were ∼70–85%, H values were ∼60% and F_r values were ∼10% for both storage conditions. The level of histamine exceeded the legal limit (5 mg/100 g fish) in eel stored without ice after 6–7 days and, in ice, after 13–14 days of storage, at which time eels were rejected by the sensory panel. The concentrations of biogenic amines were higher in eel stored in boxes without ice than in eel kept in ice. The levels of histamine in the muscle of eel kept in boxes without ice and in ice increased to the maximum levels of 17.9 mg/100 g on day 12 and 12.6 mg/100 g on day 19, respectively.     

Effect of temperature on the development of the yam moth, Dasyses rugosella Stainton (Lepidoptera: Tineidae)

Studies on the effect of temperature on the development of the yam moth, Dasyses rugosella Stainton were carried out in the laboratory at four different temperatures, 20°C, 24°C, 29°C and 33°C. The mean fecundity per female at 20°C, 24°C, 29°C and 33°C was 37.6±1.9, 84.2±2.7, 112.3±3.9 and 109.3±3.9 eggs, respectively. Oviposition period ranged from 4.7 days at 20°C to 2.9 days at 33°C. Hatchability of eggs was highest at 29°C with 70.0% and lowest at 20°C with 15.0%. The developmental time decreased with increase in temperature. The mean developmental time at 20°C, 24°C, 29°C and 33°C was 13.8±0.3, 7.8±0.2, 5.1±0.2 and 3.9±0.2 days for the egg, 90.0±0.7, 50.1±0.7, 36.1±0.8 and 23.6±0.8 days for the larval stages, 22.3±2.6, 12.2±0.2, 10.1±0.1, and 7.8±0.2 days for the pupa and 126.1±0.8, 70.1±0.8, 51.3±0.9, and 35.1±0.8 days for the period from egg hatching to adult emergence, respectively. Since fecundity, oviposition and development of the moth were impaired at 20°C, storage of yam tubers at low temperatures (well below 20°C but not lower than 12°C to avoid chilling injury) will significantly retard the development of D. rugosella and help in its control. Unmated individuals lived longer than their mated counterparts. Adult females were always larger than the males. The wing span of the female ranges from 16.9-18.5 and in males from 13.0-15.0 mm. The body length for males ranged from 5.0-6.5 and was 6.5-9.0 mm for females.     

Effects of light,temperature and water activity on the kinetics of lipoxidation in almond-based products

Lipoxidation in almond-derived products was investigated using the chemiluminescence (CL) and thiobarbituric acid-reactive substances (TBARS) methods to detect the first and later reaction products, respectively. The effects of light during storage at 5 °C, 22 °C and 40 °C were studied, as well as the effects of combined heat/water activity treatments in the 60–120 °C and 0.38–0.72 range. During storage, light was found to enhance the CL and TBARS values, and specific responses were observed in almond paste and the final Calisson product. During the heating of almond paste, as the initial water activity (a_w) increased, the CL rate constants increased during heating to 60 °C and 80 °C, but interestingly, these values decreased during further heating to 120 °C, whereas the maximum TBARS rate constants occurred at a_w 0.57 at all the heating temperatures tested. The activation energies, based on the CL and TBARS values, decreased specifically when the a_w increased from 0.38 to 0.72, giving overall values ranging from110 kJ mol⁻¹ to 60 kJ mol⁻¹. Likewise, in the same water activity range, the temperature-dependent rate constant enhancing factor (Q₁₀) decreased from 3.3 to 1.6.     

Protective effects of a synthetic soybean isoflavone against oxidative damage in chick skeletal muscle cells

The antioxidant and protective properties of a synthetic soybean isoflavone (SI) were investigated using chick skeletal (leg) muscle cells (SMC) isolated from 20-day-old Linnan yellow broiler chick embryo. Skeletal muscle cells were cultured in Dulbecco’s modified Eagle’s medium treated with 0, 12.5, 25, 50, 75 and 100 μM SI, respectively, under 80 μM H₂O₂/FeSO₄ conditions. After 24 h of incubation, SI reduced the loss of SMC under oxidative stress by H₂O₂/FeSO₄. The addition of SI significantly promoted SMC proliferation (P < 0.01). Upon treatment with 25, 50, 75 and 100 μM SI, the activity of total superoxide dismutase in the supernatant of the media was enhanced by 17.0%, 13.0%, 13.3% and 11.9%, respectively (P < 0.05). Compared to the control, the activity of glutathione peroxidase was significantly increased only at 25 μM concentration of SI (P < 0.05), and the increment was 90.7%. The activity of catalase was increased by 49.2% and 49.1%, respectively, at 75 and 100 μM SI (P < 0.01). The concentration of creatine kinase in the media was decreased by 61.6% and 60.6%, respectively, at 75 and 100 μM SI (P < 0.01). The addition of SI did not affect the activity of lactate dehydrogenase in the media. In conclusion, the SI protected skeletal muscle cells from oxidative damage, attributed to its antioxidant activity.     

Mineral content of tropical fruits and unconventional foods of the Andes and the rain forest of Colombia

A total of 68 species of starchy foods, tropical fruits, leaves and tubers (101 samples), were collected on the foothills of the Colombian Andes and in the rain forests of the Colombian Pacific coast. Their edible portion was analyzed for mineral content (Ca, P, Mg, K, Na, Cl, S, Mn, Zn, Fe, Cu, Se, Co, Ni). The foods were generally high in K (36–1.782 mg K/100 g edible portion) and low in sodium (<45 mg Na/100 g edible portion). The tree foliages had the highest contents in most of the elements, especially in calcium (280–1242 mg Ca/100 g edible portion, i.e., up to 62 g Ca/kg dry matter) and iron (0.7–8.4 mg Fe/100 g edible portion). Correlations (P < 0.001) were observed between total ash and many elements, especially Ca and Mg (r =  0.77 and 0.73, respectively). High correlations were also obtained between Ca and Mg (r = 0.93).     

Crude glycerine inclusion in Limousin bull diets: Animal performance,carcass characteristics and meat quality

Three hundred and six Limousin young bulls (7 ± 1 months of age, initial body weight 273 ± 43 kg) were used to evaluate the effect of crude glycerine supplementation on animal performance, carcass characteristics and meat quality. Animals were assigned to three different diets: Control (0% of crude glycerine), G2 and G4 (2 and 4% of crude glycerine, respectively). The diets were administrated ad libitum for 240 days (final body weight 644 ± 24 kg). Average daily weight gain, average daily feed intake, the gain:feed ratio, ultrasound measures in vivo, carcass characteristics, pH, water holding capacity, drip losses, and cooking losses were not affected (P > 0.05) by diets. Diet decreased C16:0 (P < 0.01) and C16:1 (P < 0.05) contents in meat. The G4 meat showed lower C12:0, C14:0, C17:0, C18:0, C18:1, C18:2, C18:3, c9,t11-c18:2, C20:0 and C20:4 levels (P < 0.05) than control. Glycerine increased desirable fatty acid percentages (P < 0.05) in intramuscular fat.     

Effect of multiple factors on accumulation of nucleosides and bases in Cordyceps militaris

In order to improve the accumulation of valuable metabolites (3′-deoxyadenosine, adenosine, guanosine, cytidine, uridine, adenine and uracil) in Cordyceps militaris, multiple factors such as carbon sources, nitrogen sources, metal ions and fermentation duration were investigated in liquid shake flasks. Using both potato broth and glucose as carbon sources were found to facilitate the production of nucleosides and bases most, and 2% of glucose was the most suitable initial level. Mixture of 0.3% yeast extract with 0.3% peptone was the best selection of nitrogen sources and supplementing with 0.1 mmol/L Mn²⁺ achieved the maximum biomass and biosynthesis of nucleosides and bases among all investigated metal ions. The most suitable harvest time for C. militaris was found to be the 6th day, because the production of 3′-deoxyadenosine, adenosine and uridine began to speed up from the later exponential growth phase until the 6th day. Finally, under optimal culture conditions, the contents of 3′-deoxyadenosine, adenosine, guanosine, cytidine, uridine, adenine and uracil were increased to 0.212 ± 0.014 mg/g, 5.05 ± 0.31 mg/g, 4.03 ± 0.30 mg/g, 0.556 ± 0.029 mg/g, 6.39 ± 0.33 mg/g, 0.208 ± 0.016 mg/g and 0.437 ± 0.027 mg/g, respectively.