3株小球藻对亚硒酸钠的生理响应及富硒效果的研究

小球藻是无机硒有机化的良好载体。为筛选出一株高效富硒的小球藻藻株,系统性研究了不同亚硒酸钠浓度(0～40 mg/L)对蛋白核小球藻(Chlorella pyrenoidosa)、椭圆小球藻(Chlorella ellipsoidea)和凯氏拟小球藻(Parachlorella kessleri)的生长和富硒转化效率的影响。结果显示,3株小球藻生物量和光合色素含量均随着亚硒酸钠浓度的增加而下降,但高硒浓度却有利于藻体总硒和有机硒产量的提高;其中,在相同亚硒酸钠浓度的处理下,凯氏拟小球藻的生物质增长量和光合色素含量都要高于其他2株小球藻,而且具有更高的富硒能力,该藻在8 mg/L亚硒酸钠条件下获得最高有机硒产量为404.12μg/L,有机硒比率高达90.16%,甚至在20 mg/L亚硒酸钠的高硒条件下,仍然维持生物量的正增长。综上所述,凯氏拟小球藻是一株优质的富硒藻株,具有良好的开发利用价值。     

酒精沼气双发酵偶联工艺中沼液有机酸对酒精发酵的影响

木薯酒精蒸馏废液经厌氧沼气发酵后的沼液全部回用作为酒精发酵配料的工艺用水,形成酒精沼气双发酵偶联工艺。研究发现,乙酸和丙酸是抑制酒精发酵的主要的小分子有机酸。乙酸对酒精发酵产生抑制的有效质量浓度为0.4g/dL,丙酸对酒精发酵产生抑制的有效质量浓度为0.1g/dL,乳酸质量浓度达到2.5g/dL时才会对酒精发酵产生影响。     

基于抗生素敏感性构建索罗金小球藻异养无菌体系的研究

为构建索罗金小球藻（Chlorella sorokiniana FZU60）的异养无菌体系,对其共生菌进行分离和鉴定,并考察了8种常见抗生素（头孢噻肟、卡那霉素、新霉素、氨苄青霉素、链霉素、四环素、氯霉素和红霉素）对共生菌和藻株FZU60生长的影响。结果表明：异养体系中主要的共生菌为芽孢杆菌（Bacillus）(78.76%)和甲基杆菌（Methylobacterium）(3.14%),且10 mg/L头孢噻肟、10 mg/L四环素、100 mg/L卡那霉素和100 mg/L红霉素对这两株共生菌均具有较好抑制效果,但对藻株FZU60的生长无明显影响。在此基础上,进一步考察了不同抗生素组合对藻株FZU60异养共生菌的抑制效果,发现采用10 mg/L头孢噻肟、10 mg/L四环素和100 mg/L卡那霉素的抗生素组合可成功构建异养无菌体系,且经3次传代培养后藻株FZU60仍可维持稳定的生长及无菌状态。本实验结果可为索罗金小球藻高密度异养中无菌藻种的制备提供一定的理论参考和实验依据。     

培养条件对蛋白核小球藻生长及油脂合成的影响

以富油蛋白核小球藻为出发藻株,研究自养、异养和混养培养模式对小球藻生物量和油脂含量的影响,以及异养发酵培养基葡萄糖质量浓度、氮源种类及质量浓度对小球藻生长的影响。结果表明,与自养和混养培养模式相比,采用异养发酵方式培养蛋白核小球藻可获得最大的生物量和油脂含量。通过气相色谱法测得异养蛋白核小球藻油主要脂肪酸为棕榈酸(36.07%)、油酸(34.26%)、亚油酸(20.17%)和亚麻酸(6.12%)。经单因素试验优化得到最适蛋白核小球藻生长异养发酵培养基的葡萄糖质量浓度为60 g/L,最适的氮源为酵母粉,质量浓度为4 g/L,在此条件下经192 h发酵,蛋白核小球藻生物量可达12.43 g/L,油脂产量为5.45 g/L。研究结果表明,异养发酵培养获得的蛋白核小球藻油是一种潜在且可再生的新油源。     

抑菌剂在异养小球藻发酵过程中的应用

通过抑菌圈试验、摇瓶发酵试验和50 L发酵试验,研究了安菌素、克菌灵、青霉素3种抑菌剂在小球藻培养过程中的应用,考察了抑菌剂添加对藻细胞干重和葡萄糖浓度的影响。结论:在小球藻发酵染菌4 d后,加入100 mg/L克菌灵可以有效抑制杂菌,并且不会对小球藻发酵造成影响。相同发酵周期内,小球藻染菌后抑菌发酵与正常发酵相比,藻细胞干重质量浓度达到75.2 g/L,平均干重增加速率为6.26 g/(L·d),与正常发酵相比,干重质量浓度与平均干重增加速率降低14.93%,能够成功地挽救染菌的发酵过程。而加入100 mg/L安菌素和青霉素均对小球藻的正常生长产生了抑制作用。     

酸浓度和pH值对浓香型大曲酯化酶催化活力的影响

研究了己酸、乳酸、丁酸、乙酸的浓度以及pH值对浓香型大曲酯化酶催化合成四种酯（己酸乙酯、乳酸乙酯、丁酸乙酯、乙酸乙酯）的影响。结果表明,大曲酯化酶催化合成四种酯的最适酸质量浓度分别为己酸8 g/L,乳酸1 g/L,丁酸9 g/L,乙酸12 g/L;在同一酸浓度条件下,大曲酯化酶催化合成己酸乙酯、丁酸乙酯、乙酸乙酯最适pH值均为4.5,乳酸乙酯最适pH值为6.0;大曲酯化酶催化相同物质的量浓度混合酸的试验结果表明,己酸乙酯的合成以酯化酶催化反应为主,而乳酸乙酯和丁酸乙酯的合成以化学反应为主。     

梭状芽抱杆菌L—Ⅱ基质范围的代谢研究

对梭状芽孢杆菌L—Ⅱ的含碳基质范围的研究表明：该菌株能利用乙酸、丙酸、丁酸、乙醇和丙醇。以乙醇和乙酸为底物时，主要产物是丁酸和己酸。以乙醇和丙酸为底物时，主要产物是戊酸和庚酸。以乙酸和丙醇为底物时，主要产物是丙酸、戊酸、丁酸和己酸。该菌的丙酸生物合成途径与克氏梭菌基本一致，适用于科研和浓香型酒生产。（益平）     

利用流式细胞仪筛选紫外诱变高含油小球藻

为筛选得到高含油的小球藻藻株,对若夫小球藻(Chlorella zofingensis)进行紫外诱变,尼罗红染色后再利用流式细胞仪对高含油藻株进行筛选。结果表明:紫外诱变40 min后,经流式细胞仪筛选,分选出的90%的突变藻株总脂含量都高于野生对照株。最终筛选到的藻株S16经培养总脂产量达到2.4 g/L,比野生对照株提高了50%。     

硒浓度对小球藻生长、生物富集的影响

研究了硒浓度对小球藻生长的影响,以及硒浓度对小球藻生物富集的影响,并对相关的可能机理进行了讨论.研究发现,高浓度的硒对小球藻生长具有抑制作用,当硒浓度超过1mg/L时会产生抑制作用;当硒浓度增至12mg/L时,发现藻细胞生长几乎停止.小球藻对硒的生物富集随着硒浓度的增加而增加,硒浓度为1mg/L时,藻体的硒含量为9.92μg/g;当硒浓度为8mg/L时,藻体的硒含量达到了83.66μg/g.小球藻富硒培养较适宜的硒添加浓度是4mg/L～8mg/L.     

甘蔗制糖文摘(三)

高温／中温两相厌氧消化反应中有机酸的变化──赵庆良等，环境科学．1996（3），44～47由于有机酸是厌氧消化反应过程中有机物分解时产生的重要中间产物，大部分甲烷气都是由有机酸的进一步分解而产生的，因此本大着重探讨有机酸的变化情况。试验结果表明：混合基质经高温消化后择没有机酸含量有所增加，平均在3346～5529mg／L，进一步经中温消化且系统达到稳定后其含量平均为43～433。g／L；乙酸、丙酸、丁酸和成酸是进料混合基质和高温消化后基质中主要的有机酸种类。固定化微生物治理味精废液──敬一兵，环境工程，1996（3），52～53…     

利用甲醇厂CO_2尾气培养小球藻

利用甲醇厂所排放的高浓度CO2尾气培养小球藻,研究不同的通气方式、培养方式对小球藻生长的影响。实验结果表明,当通气速率为300 mL/min、CO2浓度为10%并以10 min/h的间歇通气方式培养时能够提高小球藻的生物量,生物量和产率分别为0.681 g/L和0.082 g/(L.d)。对氮源和磷源采用补料的方式进行培养时对小球藻生长具有促进作用,最高生物量和产率分别达到0.789 g/L和0.088 g/(L.d)。当培养基的更新率为20%时能够获得较高的生物量。利用醋酸进行兼性培养时,一次性添加50～125μL/L醋酸都可促进小球藻生长,每日添加10～25μL/L的醋酸时,可明显促进小球藻生长,最高生物量和产率分别为0.933 g/L和0.111 g/(L.d),分别是空气对照的1.3倍和1.4倍。说明利用甲醇厂高浓度CO2尾气培养小球藻提高生物量是可行的。     

有机酸含量对葡萄酒发酵的影响

为了考察有机酸含量对葡萄酒发酵的影响,在初始有机酸含量分别为4 g/L、7 g/L和10 g/L的模拟葡萄汁中接种酵母,于25℃发酵,比较不同初始有机酸含量对葡萄酒发酵过程中酵母生长、降糖、有机酸及挥发性化合物含量的影响。结果表明,初始有机酸含量7 g/L时酵母生长和耗糖最慢,乙醇和乙醛产生量最低;随初始有机酸含量升高,pH值、酒石酸、苹果酸、乳酸、乙酸、总酸、乙酸乙酯和总酯含量呈下降趋势;琥珀酸、正丙醇、异丁醇、异戊醇及总高级醇含量呈上升趋势;但对β-苯乙醇、正丁醇及丁酸乙酯含量影响较小。初始有机酸含量10 g/L的葡萄酒发酵过程酵母生长快、降糖最快,产酸含量适中,同时香气较好。因此,葡萄酒发酵的适宜初始有机酸含量为10 g/L。     

Green technology for conversion of food scraps to biodegradable thermoplastic polyhydroxyalkanoates

A new technology is developed and demonstrated that couples anaerobic digestion of food scraps with production of biodegradable thermoplastics, polyhydroxyalkanoates (PHAs). The food wastes were digested in an anaerobic reactor producing four major organic acids. The concentrations of acetic, propionic, butyric, and lactic acids reached 5.5, 1.8, 27.4, and 32.7 g/L, respectively. The fermentative acids were transferred through membranes via molecule diffusion into an air-bubbling reactor where the acids were utilized to produce PHAs in an enriched culture of Ralstonia eutropha. With a silicone rubber membrane, butyric acid and small amounts of acetic and propionic acids were transferred and used, producing a homopolymer PHA, poly(3-hydroxybutyrate). The dry cell weight and PHA content reached 11.3 g/L and 60.2% (w/w), respectively. With a dialysis membrane, the mass transfer rates of fermentative acids were enhanced, and the PHA production was significantly improved. The dry cell weight and its PHA content reached 22.7 g/L and 72.6% (w/w), respectively. The formed PHA was a copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate (HV) with 2.8 mol % HV monomer unit. The polymer content (72.6% of dry cell mass) reported in this study is the highest one obtained from organic wastes and is comparable with the PHA content from pure glucose fermentation.     

Optimization of Growth of Lactobacillus casei ASCC 292 and Production of Organic Acids in the Presence of Fructooligosaccharide and Maltodextrin

ABSTRACT: Growth of Lactobacillus casei ASCC 292 was optimized in the presence of 2 prebiotics, namely fructooligosaccharide (FOS) and maltodextrin, and the patterns of organic acids produced was monitored. Perturbation plot, response surface, and coefficient estimates showed that inoculum size and concentration of prebiotics had significant quadratic effects on growth, with FOS showing the most prominent quadratic change. The second-order polynomial regression model estimated the optimum inoculum size and the concentrations of FOS and maltodextrin for the growth of L.casei ASCC 292 to be 1.67% w/v, 4.82% w/v, and 6.93% w/v, respectively.Validation experiments showed that the predicted optimum condition was better than the high and low levels of the factors, and at center points. Response surface methodology (RSM) was reliable in developing a model, optimizing factors, and analyzing interaction effects. Analysis of individual organic acids production by L. casei ASCC 292 showed that highest amount of lactic acid was produced, followed by propionic, acetic, formic, and butyric acid. The production of lactic acid was greatly influenced by the inoculum size, and the organism was relatively sensitive to the end-product of maltodextrin fermentation. Production of acetic acid was greatly influenced by the inoculum size and was inhibited by end-product of the maltodextrin fermentation. Increased concentration of FOS contributed to the increased production of propionic acid. The response surface of butyric and formic acid showed that the production of these acids were growth associated.     

窖泥中高产丁酸梭菌的筛选、鉴定及其生长耐受性研究

从浓香型白酒窖泥中通过传统微生物分离手段筛选高产丁酸的梭菌,并对分离菌株进行形态学观察、分子生物学鉴定及生长耐受性分析研究。结果表明,通过发酵产物检测获得两株高产丁酸菌株,编号为AB2RH027和AB2RH033,经过菌落观察、革兰氏染色及分子生物学鉴定两株菌均为丁酸梭菌（Clostridium butyricum）。通过耐受性实验,确定菌株AB2RH027最适生长pH值为8,最适生长温度为40 ℃,酒精耐受性为15%vol,发酵24 h丁酸产量为5.29 g/L;菌株AB2RH033最适生长pH值为8,最适生长温度为35 ℃,酒精耐受性为18%vol,发酵24 h丁酸产量为5.23 g/L。该研究筛选得到的两株高产丁酸的菌株在白酒品质提升方面具有一定应用价值。     

高耐受性醋酸菌的筛选及发酵特性研究

从工业醋醅中分离出5株耐乙醇、耐高温的产醋酸菌株,利用生理生化试验和16S rDNA同源序列分析,初步认定其为巴氏醋杆菌（Acetobacter pasteurianus）。通过对其耐乙醇、耐高温发酵特性的研究,发现菌株FY4可耐受体积分数为12%的乙醇和43 ℃高温,在30 ℃和体积分数10%乙醇条件下产酸量达到最高,为61.2 g/L。在10 L发酵罐试验中,菌株FY4的产酸量始终高于醋酸菌AS1.41,在37 ℃和体积分数10%乙醇条件下,菌株FY4产酸量达到42.2 g/L,而在此条件下工业菌株AS1.41几乎停止产酸。结果表明菌株FY4具有高耐受性,可产生大量的醋酸,在工业生产中具有潜在应用价值。     

IMPROVED METHODS FOR DETERMINATION OF CERTAIN ORGANIC ACIDS IN PASTEURIZED AND UNPASTEURIZED LIQUID AND FROZEN WHOLE EGG

ABSTRACT— This work was supported in part by National Institutes of Health Contract Ul-00131-06.
Liquid whole egg to which acetic, butyric, lactic, propionic and succinic acids were added, or which had previously been incubated at room temperature to different microbial populations, was pasteurized at 60.5°C for 3.5 min. A liquid-liquid extraction method, including the centrifugation of the denatured egg, was developed for extraction of all 5 acids. The recovery of the 5 acids was evaluated by an improved gas-liquid chromatographic procedure. Lactic and succinic acids were recovered from the liquid whole egg samples and chromatographed as their butyl ester derivatives and acetic, propionic and butyric acids were recovered and chromatographed as the acids per se. The fresh egg samples containing small amounts of acetic and lactic acid and liquid whole egg product, incubated until a microbial population of 4.5 × 10⁶ was obtained, con tained only these 2 acids at a higher concentration. The pasteurization process did not affect concentration of the added short-chain organic acids or those which accumulated during the incubation period; thus, accurate and valid analyses of the short-chain organic acids can be accomplished in pasteurized whole egg products.     

Organic acids and volatile flavor components evolved during refrigerated storage of kefir

Kefir samples were prepared and transferred to sterile jars for storage at 4 degrees C. After 0, 7, 14, and 21 d of storage, the pH, organic acid, and volatile flavor component content were determined to monitor possible flavor changes during storage. Stored samples were analyzed for organic acid (orotic, citric, pyruvic, lactic, uric, acetic, propionic, butyric, and hippuric) content by HPLC with UV detection at 275 nm. Acetoin, ethanol, acetaldehyde, and diacetyl were monitored using gas chromatography equipped with a headspace autosampler. There was no significant decrease in average pH of samples between d 0 and 21 of storage (P>0.05). Lactic acid concentration increased during storage, reaching a maximum of 7739 ppm by d 21. Orotic and citric acids increased slightly during storage. Although pyruvic and hippuric acids are produced during fermentation, neither was detected during storage. Acetic, propionic, and butyric acids were not detected during kefir storage. Ethanol concentrations increased during storage and reached 0.08% by d 21. The amounts of acetaldehyde and acetoin, common flavor substances in many cultured dairy products, increased during fermentation. Acetaldehyde content in kefir samples doubled from d 0 to 21, reaching a final concentration of 11 microg/g. During storage, the concentration of acetoin decreased from 25 ppm on d 0 to 16 ppm on d 21. However, diacetyl, another common flavor component in cultured dairy products, was not detected during fermentation or storage.     

以废弃毕赤酵母为高效氮源强化丁酸发酵生产

为有效利用固态废弃毕赤酵母,高效发酵生产丁酸,建立了以80 g/L玉米淀粉为基础培养基、以废弃毕赤酵母处理液为高效有机氮源,连续补加葡萄糖的丁酸发酵工艺。7 L厌氧发酵罐下,发酵20h,先以1∶4的比例添加250~400 g/L规格的废弃酵母处理液替代昂贵的发酵用复合培养基,再连续补加葡萄糖浓缩液。最终的丁酸质量浓度为45 g/L、得率为40%,丁酸占总酸的比率(butyric acid ratio over total organic acids,B/TA)≥0.90的较高水平。该工艺可以有效地利用来自于废弃酵母处理液中的氨基酸,提高细胞生长速度和浓度,增强丁酸合成所依赖的NADH再生速度,间接改善了丁酸发酵性能。与此同时,实现了废弃生物质的有效利用和资源化。