影响被孢霉产生含r—亚麻酸油脂的几种因素

对培养条件,如氮源种类、Ｃ／Ｎ比率、ｐＨ、种子种龄与接种量对被孢霉（Ｍｏｒｔｉｅｒｅｌｌａ ｓｐ）Ｍ１４菌株的细胞生长和油脂形成的影响作了研究。结果表明,二级发酵优于一级发酵。Ｍ１４菌株二级发酵时,种子种龄以４９ｈ为适,接种量宜大（３０％）。无机氮源有利于不饱和脂肪酸的产生,有机氮源有利于细胞的增殖。低Ｃ／Ｎ比率有利于菌丝体产量的提高,提高Ｃ／Ｎ比率则能促进菌体细胞内的油脂合成。ｐＨ对菌体生长、油     

深黄被孢霉高产脂变株的选育及其发酵的研究

以深黄被孢霉（Mortierellaisabellina）AS3．3410为出发菌株,经紫外线,硫酸二乙酯和亚硝基胍复合诱变处理,选育成功高产脂深黄被抱霉M018变株,其摇瓶培养菌体油脂含量达65．6％,比出发菌株提高133％。60m³罐三级发酵培养菌体油脂含量高达79．2％,生物量达37．8g／L．气相色谱分析表明变株M018r-亚麻酸的含量比出发菌株提高了53％。连续传代试验表明M018是一稳定的变株。该变株油脂合成的最佳碳源为葡萄糖,最佳氮源为酵母膏,最佳C／N为     

丝孢酵母高甲硫氨酸突变株的选育及营养调控

以丝孢酵母(Trichosporon Behr)ST851为原始菌株,经紫外线诱变,在含乙硫氨酸的双层平板上筛选到多株抗乙硫氨酸突变株。其中ST851-10株抗乙硫氨酸浓度达到350μg/ml,其菌体蛋白质含量由40.5％提高到44.3％,菌体甲硫氨酸含量由20.45mg/g-DCW增加到29.32mg/g-DCW。在以苹果渣为碳源、尿素为氮源、硫酸镁作硫源的最适培养条件下,固态发酵24h后,蛋白质和甲硫氨酸含量较原始菌株分别提高了15.8％和44.9％。培养基中C/N值低有利于甲硫氨酸的合成,C/N值高则适合于菌体生长。在苹果渣固态发酵过程中,适当补加氮源既有利于菌体生长和甲硫氨酸的合成,又可起到调节培养基pH值的作用。     

丝状真菌被孢霉产生油脂的研究

被孢霉的三个菌株Mortierella sp.M10,M13与M14生长在以葡萄糖为碳源、尿素为氮源的液体培养基中,所得到的菌丝体内堆积含γ-亚麻酸的油脂。油脂产率以M10菌株为高,而油脂中的γ-亚麻酸含量却以M14菌株为高。这种油脂的脂肪酸中,所含饱和脂肪酸主要有豆蔻酸(C14:0),棕榈酸(C16:0)和硬脂酸(C18:0);所含不饱和脂肪酸主要有棕榈油酸(C16:1),油酸(C18:1),亚油酸(C18:2)以及γ-亚麻酸(C18:3,n-6)。上述被孢霉菌株的培养物接种在含葡萄糖、尿素的培养基中生长大约48小时后,菌丝体顶端细胞形成鼓胀球状,此后仍继续胀大。菌体细胞形态的这种特异变化,可能与胞内油脂的累积有关。     

氨水中和Lactobacillus delbrueckiisubsp. lactis BME5-18M发酵生成L-乳酸铵的研究*

实验确定了Lactobacillusdelbrueckiisubsp lactisBME5-18M接种的最佳种龄为24h。以氨水取代传统的中和剂碳酸钙中和发酵生成的乳酸、调控发酵液的pH ,考察了不同pH值对菌体生长和产酸的影响 ,确定了菌种生长和产酸的较适pH值为 6.5。考察了底物流加速度对菌种生长和产酸的影响 ,对间歇和流加发酵时菌体的生长量和产酸量进行了动力学关联。在较适pH值 6.5和较佳流加速度 25mL/h条件下 ,乳酸的产量可达到 136.8g/L ,产率为1.     

解脂假丝酵母利用正癸烷产生癸二酸的研究

我们从石油酵母和细菌中,以及采自不同地区的油浸土、植物的花和果实中,经过了广泛筛选,得到能利用正癸烷产生癸二酸的优良菌株酵母19—2。经鉴定为解脂假丝酵母(Candidalipotytica)。初步研究了该菌在摇瓶发酵中产生癸二酸的条件,在正癸烷、尿素、磷酸二氢钾、玉米浆和醋酸钠组成的培养基上,可以氧化正癸烷而产生癸二酸。发酵96小时左右,投油比10％(v／v),发酵液中癸二酸含量为32—40克／升,对正癸烷收率为43—55％。在发酵培养中,用醋酸钠作碳源,菌生长良好。正癸烷不作唯一生长碳源,只由菌体酶催化转化为癸二酸。产酸的最适氮源是尿素,玉米浆对产癸二酸较为有利。在普通300,~y{---角瓶中,装液量对产癸二酸影响很大。在带有挡板三角瓶中,装量对产癸二酸亦有关系。此外,种子种龄、种子生长的最终PH、接种置对产癸二酸亦有很大影响。并做了500升罐的扩大试验,产酸稳定,重现性良好。     

蛭弧菌J26转化山梨糖生产2—酮基—L—古龙酸发酵条件的研究

对新分离得到的能产生维生素C前体2-酮基-L-古龙酸（2-KGA）的优良菌株蛭弧菌J26,进行了一系列摇瓶发酵条件试验,通过添加一些其它有机碳源或其它氮源,可使蛭弧菌J26摇瓶发酵产生2－KGA由50-60mg／ml上升到70mg／ml以上,采用SMA探针技术的流加发酵摇瓶试验产生2－KGA可以达到83．9－91．7mg／ml。本文还对发酵时种子液的接种量、发酵初始精浓度、初始pH值等对产酸的影响,进行了比较研究,对摇瓶发酵全过程的单一菌体形态特征进行了电子显微镜观察。     

中度嗜盐菌产木聚糖酶发酵条件的研究

中度嗜盐菌在盐碱环境下生长繁殖,其产生的木聚糖酶也同样具有在盐碱环境下发挥作用的特性。本文对一株中度嗜盐菌的产木聚糖酶活性进行了初步研究。研究包括氮源、液体种子接种量、培养温度、pH值、培养时间等因素对该菌株产木聚糖酶能力的影响。结果表明,最佳培养氮源为蛋白胨;最佳产生木聚糖酶的发酵条件是液体种子接种量为6%,温度为35℃,pH值7,培养时间为4 d。     

真菌发酵生产EPA及DHA影响因素的研究进展

对真菌发酵生产EPA及DHA的影响因素进行综述 ,介绍了菌种、碳源、氮源、C/N比、pH值、温度、发酵时间、通气量、代谢途径的调控、种龄和接种量等因素对EPA及DHA产量的影响。     

培养幽门螺旋杆菌的发酵工艺研究

采用摇瓶微需氧培养技术,模拟发酵条件,探讨不同因素（pH、转速、种子菌接种量）对幽门螺旋杆菌（Helicobacter pylori,Hp）生长的影响,优化工艺参数,并级联放大到10L发酵罐发酵,经多次实验,建立了稳定的Hp发酵工艺,24h发酵细菌的最大吸光度A600达3.89,收获菌体湿重达5.2g／L。     

培养条件对产油微生物生长的影响

为了筛选出高产油菌株, 首先采用细胞形态学方法与细胞化学方法(苏丹III染色法)对4株高产油脂菌株进行初筛, 并通过索氏提取法对初筛菌株油脂含量进行分析, 确定M2菌株为实验菌株, 其油脂含量达53.09%。为了增加产油微生物油脂产量, 本试验考察了不同发酵条件对其细胞生长和油脂积累的影响。优化工艺参数为: 10° Bx玉米皮渣水解液为培养基质, 0.2% NaNO3为氮源, pH 6.0、28oC下发酵培养6 d, 微生物油脂含量75.21%, 菌体生物量30.40 g/L, 油脂产量22.86 g/L。气相色谱分析表明该油脂的脂肪酸组成与植物油相似, 主要含有16碳和18碳系脂肪酸, 可作为生物柴油的原料, 不饱和脂肪酸含量达68%, 可应用于医药化工领域。     

Efficient concomitant production of lipids and carotenoids by oleaginous red yeast <Emphasis Type="Italic">Rhodotorula glutinis</Emphasis> cultured in palm oil mill effluent and application of lipids for biodiesel production

Rhodotorula glutinis TISTR 5159 is oleaginous red yeast that accumulates both lipids and carotenoids. It was cultured in palm oil mill effluent (POME) with only the addition of ammonium sulfate and Tween 20 as a suitable nitrogen source and surfactant, respectively. Response surface methodology (RSM) was applied to optimize initial chemical oxygen demand (COD) in POME, C/N ratio, and Tween 20 concentration for concomitant production of lipids and carotenoids. Among three investigated factors, C/N ratio contributed a significant effect upon lipid and carotenoids production. Analysis of response surface plots revealed that the optimum C/N ratio for the biomass was 140, while that for lipid content and carotenoids were higher at 180 and 170, respectively. The high level of the nitrogen source (with a low C/N ratio) enhanced the biomass, making the accumulation of lipids and carotenoids less preferable. Hence, the two-stage process was attempted as an optimal way for cell growth in the first stage and product accumulation in the second stage. The lipid yield and carotenoid production obtained in the two-stage process were higher than those in the one-stage process. In the semi-continuous fermentation, R. glutinis TISTR 5159 accumulated high lipid content and produced a considerably high concentration of carotenoids during long-term cultivation. Additionally, efficient COD removal by R. glutinis TISTR 5159 was observed. The biodiesel produced from yeast lipids was composed mainly of oleic and palmitic acids, similar to those from plant oil.     

氮源及碳氮比对产朊假丝酵母合成谷胱甘肽的影响

研究了N源对产朊假丝酵母细胞生长和谷胱甘肽(GSH)合成的影响。在此基础上,分别以(NH4)2SO4和尿素作为单一N源,摇瓶条件下研究了不同C、N比对GSH发酵的影响。结果发现尿素有利于细胞生长,而(NH4)2SO4更有利于GSH的合成,并且酵母细胞在利用这2种N源合成GSH时,各自具有最佳的C、N比((NH4)2SO4为8.3 mol/mol,尿素为5.6 mol/mol)。最佳C、N比下的GSH分批发酵结果显示,尿素是更合适的N源,最终细胞干质量和GSH产量可以分别达到16.48 g/L和246.4 mg/L。最后分别采用发酵动力学模型和代谢网络分析对该结果产生的原因进行了定量解释。     

Improving lipid production from bagasse hydrolysate with Trichosporon fermentans by response surface methodology

Oleaginous yeast Trichosporon fermentans was proved to be able to use sulphuric acid-treated sugar cane bagasse hydrolysate as substrate to grow and accumulate lipid. Activated charcoal was shown as effective as the more expensive resin Amberlite XAD-4 for removing the inhibitors from the hydrolysate. To further improve the lipid production, response surface methodology (RSM) was used and a 3-level 4-factor Box-Behnken design was adopted to evaluate the effects of C/N ratio, inoculum concentration, initial pH and fermentation time on the cell growth and lipid accumulation of T. fermentans. Under the optimum conditions (C/N ratio 165, inoculum concentration 11%, initial pH 7.6 and fermentation time 9 days), a lipid concentration of 15.8g/L, which is quite close to the predicted value of 15.6g/L, could be achieved after cultivation of T. fermentans at 25°C on the pretreated bagasse hydrolysate and the corresponding lipid coefficient (lipid yield per mass of sugar, %) was 14.2. These represent a 32.8% improvement in the lipid concentration and a 21.4% increase in the lipid coefficient compared with the original values before optimization (11.9g/L and 11.7). This work further demonstrates that T. fermentans is a promising strain for lipid production and thus biodiesel preparation from abundant and inexpensive lignocellulosic materials.     

耐盐碱菌株的分离筛选及生物学特性和盐碱去除效率的研究

将采集于黑龙江省大庆市盐碱地的土壤样品经适当稀释后,涂布于高盐碱的培养基中,分离纯化获得41株菌。经过不断分别或同时提高培养基的盐、碱浓度进行耐盐碱菌株的筛选,获得1株耐盐碱菌株7,其在pH值13且盐(NaCl)浓度达到200g.L-1的培养基中仍可生长,该菌株既属于极端嗜盐微生物又属于极端嗜碱微生物。分别于不同碳源、氮源、碳氮比、初始pH值和培养温度条件下,对菌株7的生物学特性和盐、碱去除能力进行研究。结果表明,该菌株生长和去除盐碱的最佳碳源为牛肉膏;最佳氮源为蛋白胨;碳氮比为4:1时,最有利于菌株的生长和对盐、碱的去除;菌株生长的最适初始pH值为9.5,最适温度为20℃,而盐、碱去除的最佳温度则为30～35℃。     

高分子量聚谷氨酸的微生物合成及其发酵过程的研究

目的：以枯草芽孢杆菌纳豆亚种为出发菌株,考察不同碳氮源及NaCl浓度、谷氨酸、种龄、接种量对微生物发酵产1-聚谷氨酸的影响,以提高γ-聚谷氨酸的产量。方法：该菌菌种活化后,接入种子培养基,于37℃、200r／min震荡培养18h,然后按2％接种量接入不同发酵培养基进行发酵培养。γ-聚谷氨酸分离纯化后,根据其产量筛选最适发酵培养基组成及发酵条件,并对产物进行分析测定。结果：①最佳碳氮源分别为葡萄糖、蛋白胨,NaCl浓度为30g／L、种龄15h、接种量3％,且需在培养基中添加谷氨酸。②该菌株在最适条件下发酵56h时,γ-聚谷氨酸产量达32．7g／L,凝胶渗透色谱分析其相对分子质量为426kDa,呈多分子质量聚集体形式。③γ-聚谷氨酸的合成与菌体生长并非完全同步。结论：γ-聚谷氨酸作为一种天然的、可生物降解的、对环境和人体无害的多聚物,可由微生物发酵合成,且在此适宜条件下产量较高。     

响应面法优化细菌EHB01对铜绿微囊藻(Microcystis aeruginosa)的溶藻条件

【背景】水体富营养化导致的蓝藻水华对淡水资源造成了严重污染。利用环境友好型的溶藻菌可有效控制蓝藻的生长,是防治蓝藻水华形成的有效途径之一。【目的】优化溶藻细菌EHB01对铜绿微囊藻(Microcystis aeruginosa)的溶藻条件,以期为治理蓝藻水华污染提供高效的溶藻菌制剂。【方法】采用单因素试验对溶藻的发酵液浓度、温度、光照以及C:N和N:P进行分析,并对溶藻细菌EHB01发酵液的碳源、氮源和p H进行优化。基于单因素试验,选用中心组合试验设计(central composite design,CCD)确定关键因子的最佳数量水平,并以Desig-Expert 8.0.5进行回归分析,通过响应面分析获得溶藻效果最佳的参数。【结果】发酵液浓度对溶藻率的影响表现为持续上升;温度对溶藻率表现为先上升后下降;而光照、C:N和N:P均对细菌EHB01发酵液溶藻率的影响表现为先降低后上升的趋势。溶藻细菌EHB01发酵液所需的最佳碳源为蔗糖,氮源为硝酸钾,pH为7.5,优化条件下溶藻率达86.97%,与优化之前相比提高了21.72%。【结论】采用响应面法优化得出溶藻细菌EHB01发酵液最优的培养条件,且模型拟合效果较好,可为溶藻菌制剂工业化发酵提供有效依据。     

一株荧光假单胞杆菌的分离鉴定与反硝化特性

【目的】从污水厂的活性污泥中获得一株高效反硝化细菌。【方法】采用低温驯化,进行初筛、复筛选取一株反硝化活性最高的菌株,命名为L2,通过形态学、生理生化特征及16S r RNA基因序列分析研究其分类地位,系统研究理化因素对该菌株反硝化性能的影响。【结果】菌株在低温条件下能够稳定高效地进行反硝化,鉴定该菌株为荧光假单胞杆菌(Pseudomonas fluorescens),其反硝化最适接种量为10%,温度为20°C,p H为7.0,盐浓度为0.5%,碳源为葡萄糖,C/N为5.0,能够耐受较高初始硝态氮浓度。【结论】菌株L2是一株耐低温、耐高浓度初始硝态氮、耐低C/N、兼性厌氧、高效反硝化的荧光假单胞杆菌。     

Production of a novel polygalacturonic acid bioflocculant REA-11 by Corynebacterium glutamicum

The production of a novel polygalacturonic acid bioflocculant REA-11 from a newly isolated strain, Corynebacterium glutamicum CCTCC M201005, was investigated. Sucrose was chosen as a carbon source for REA-11 production. Complex nitrogen sources containing urea and an organic nitrogen compound enhanced both bacterial growth and REA-11 production, among which urea plus corn steep liquor was shown to be the most efficient combination. A cost-effective medium for REA-11 production mainly comprised 17 g/l sucrose, 0.45 g/l urea, and 5 ml/l corn steep liquor, under which conditions the flocculating activity reached 390 U/ml. The molar ratio of carbon to nitrogen (C/N) significantly affected REA-11 production, where a C/N ratio of 20:1 was shown to be the best. Interestingly, by simultaneously feeding sucrose and urea at a C/N ratio of 20:1 at 24 h of fermentation, REA-11 production (458 U/ml) was enhanced by 17% compared to the control. In a 10 l jar fermentor, lower dissolved oxygen tension was favorable for REA-11 production: a flocculating activity of 520 U/ml was achieved at a kappaLa of 100 h(-1). REA-11 raw product is relatively thermo-stable at acidic pH ranges of 3.0-6.5. Preliminary application studies showed that REA-11 had stronger flocculating activity to Kaolin clay suspension compared to chemical flocculants. In addition, the capability of decolorizing molasses wastewater indicates the industrial potential of this novel bioflocculant.     

枯草芽孢杆菌B47菌株高产抗菌物质的培养基及发酵条件优化

枯草芽孢杆菌Bacillus subtilis B47菌株为番茄内生细菌, 也是玉米小斑病拮抗菌, 能产生对玉米小斑病菌有强烈抑制作用的抗菌物质。以B47菌株发酵液的无菌滤液对玉米小斑病菌的抗菌活性为检测指标, 测定B47菌株产抗菌物质培养所需的最佳碳、氮源和无机盐, 并通过正交试验法对该菌株产抗菌物质的培养基配方和摇瓶发酵条件进行优化。研究结果表明, B47菌株产抗菌物质最佳碳、氮源和无机盐分别为蔗糖、酵母浸膏和MgSO4·7H2O, 最优培养基是YSB (Yeast extract-sucrose-beef extract)培养基, 其配方为: 蔗糖2%, 酵母浸膏2%, 牛肉浸膏1.5%, MgSO4?7H2O 0.06%, FeSO4·7H2O 0.000 9%, 最优发酵条件组合为: 30 °C, pH 7.0, 170 r/min摇床培养6 d, 接种量为1%, 装液量为40 mL/200 mL。