白腐菌Funalia trogii以稻草进行固体发酵产漆酶的研究

研究了硬毛粗毛盖孔菌Funalia trogii以稻草为培养基质进行固体发酵产生高活性漆酶。在筛选合适的碳源、氮源和硫酸铜浓度的基础上,开展四因素正交实验,筛选出最优培养方案,即以稻草粉为基质,淀粉0.5%、尿素0.3%、硫酸铜1%(质量分数),湿度为70%的条件下培养7天后漆酶活性即可达到145.42 U/gds。在最优方案条件下连续培养,发现该菌在稻草粉培养基质上培养28天产生的漆酶活性能达到最高,为1 518.76 U/gds。而在增加培养规模的情况下,以最优方案培养28天的漆酶活性平均能达到1 753.49 U/gds。     

落叶松附毛孔菌稻草液态发酵产漆酶条件优化

以木质纤维素类物质为主要基质,培养白腐真菌液态发酵产漆酶具有重要应用前景。筛选到一株漆酶高产菌,鉴定为落叶松附毛孔菌Trichaptum laricinum,优化培养条件的基础上,考察稻草和硫酸铜对该菌漆酶产量的影响。正交试验获得T.laricinum最适产漆酶条件为:麦芽浸粉为氮源,其浓度为7 g/L,硫酸铜浓度为80 mg/L,pH值4。液态发酵15 d时该菌漆酶产量达到19.2 U/mL;稻草作为主要营养基质可以显著提高该菌漆酶产量,稻草添加量为4.5 g/L时,漆酶量最大;发酵前期,无Cu~(2+)的培养环境有利于T. laricinum营养生长,在发酵中后期Cu~(2+)加入有利于漆酶分泌,第5 d加入Cu~(2+)效果最好,发酵15d时漆酶产量为23.5U/m L。T.laricinum可以利用稻草为主要基质发酵产漆酶,是一株具有潜在应用价值的漆酶工业生产菌株。     

白腐菌Coriolus versicolor的培养及产漆酶条件的研究

从液体和固体两种培养体系出发,探索不同培养条件对白腐菌分泌漆酶的影响。液体培养体系中,白腐菌的最佳生长和漆酶分泌条件是缓冲体系起始pH值为5.8、摇瓶转速为180r/min、装液量为100mL/500mL;在所有测试的碳源中,葡萄糖为最佳碳源,在二糖中以蔗糖为碳源,漆酶酶活相对较高,而多糖(如淀粉)不利于漆酶的生成。在所有测试的氮源中,硝酸铵和酒石酸铵组合为最佳氮源,氯化铵和尿素的效果次之,蛋白胨的效果最差。当碳源与氮源质量比(C/N)大于10∶1,即高碳低氮的培养条件有利于漆酶的生成。芳香化合物诱导剂ABTS浓度为0.5～1mmol/L时,对产漆酶的影响最明显,与限氮培养基中产酶相比可提高酶活5倍,约为230IU/mL;0.5mmol/L愈创木酚、1mmol/L阿魏酸和0.05mmol/L二甲基苯胺的添加可提高酶活约2～2.5倍;0.01mmol/L黎芦醇的添加可提高漆酶酶活约1.6倍。由于酪氨酸具有酚类化合物的特点,也可作为一种无毒的天然漆酶诱导剂。菌株漆酶活力在固体培养体系明显低于在液体培养体系。     

白毒鹅膏菌漆酶的诱导及其对直接黑G的脱色作用

白毒鹅膏菌是一种重要的产生胞外漆酶的菌株.探讨了漆酶在各种营养条件下的分泌情况以及对直接黑染料的降解情况.结果表明,以麸皮作为碳源,酵母膏作为氮源,在120 r/min的转速下培养,漆酶的产量最高.另外染料在较低质量浓度(10 mg/L)时,漆酶对染料的降解迅速而彻底,在pH=4.5、50℃条件下,偶氮染料直接黑G能被粗酶液(10 U/L)有效地降解,在120 r/min时降解率达到84.4%.     

黄曲霉YZC产纤维素酶的固态发酵条件优化

为了提高黄曲霉YZC固态发酵所产纤维素酶的酶活力,采用单因素试验分别研究了碳源、氮源、氮源质量浓度、培养温度、培养时间、初始p H值、接种量对纤维素酶酶活的影响。优化后的固态发酵条件为：碳源为质量比3∶2的稻草粉与麸皮,氮源为NH4Cl,其质量浓度为10 g/L,培养温度为30℃,培养时间为7 d,初始p H值为4.0,接种量为20%。在此基础上,采用响应曲面法对影响纤维素酶活的3个因素包括氮源NH4Cl的质量浓度、培养时间、初始p H值进行优化,以期提高纤维素酶酶活。通过响应曲面分析得到滤纸酶活力与这三个因素的最优回归方程,确定滤纸酶活力最大时的最佳组合为NH4Cl质量浓度为8.5 g/L、培养时间为6.5 d、初始p H值为4.5,该条件下滤纸酶活力为10.87 IU/m L,是优化前的2.39倍,并与响应曲面拟合所得方程的预测值10.50 IU/m L符合良好。     

白构菌液体发酵合成漆酶的培养条件研究

提高底物碳源的聚合度和采用有机氮源有利于诱导白构菌 (Flammulinavelutipes)合成漆酶 ,实验结果表明选择淀粉和黄豆粉作为白构菌液体发酵制取漆酶的碳源和氮源有良好的效果。采用振荡培养能够强化白构菌液体发酵体系内的传质作用 ,更适于菌体的生长及漆酶的合成。以1 80r/min速率振荡培养白构菌 1 6d ,漆酶活力可达到 3 0 8IU/mL ,比静置培养提高 83 .3 %。经活化后的白构菌接种并在振荡条件下培养可显著提高漆酶活力 ,1 80r/min振荡培养 6d后漆酶活力可达到 1 749IU/mL。     

啤酒废水培养复合型生物絮凝剂产生菌的研究

从空气和活性污泥中筛选出絮凝剂产生菌,对其进行筛选并纯化,得到絮凝活性较高且稳定的菌株M1和M2.以啤酒废水为廉价培养基,对这2株菌的复合菌(M1-2)进行培养,考察各种因素对复合菌絮凝效果的影响.结果表明,M1-2的最佳培养条件为:直接利用啤酒废水,不另外添加碳源和氮源,只需添加质量分数O.5%的KH2PO4;温度30℃,培养基初始pH=8.5,培养时间48h,摇床转速为160r·min-1.在此条件下所产生的絮凝剂对质量浓度为50g·L-1的高岭土悬液絮凝率可达95.0%,说明啤酒废水中含有丰富的营养物质,作为培养基为絮凝剂产生菌M1-2种提供丰富的碳源和氮源,从而可降低培养成本.     

直接降解木质素的漆酶/木聚糖酶体系的合成

对一株高产漆酶及伴有木聚糖酶和少量纤维素酶的菌株,用不同的碳源和氮源对其调控培养,合成漆酶/木聚糖酶体系。实验结果表明,最佳的碳源是可溶性淀粉,用它作碳源,漆酶活性高达730IU/mL,木聚糖酶活性是4.49IU/mL,纤维素酶活性只有0.23IU/mL;最佳的氮源是蛋白胨,用其作氮源,合成漆酶活性可达812IU/mL,木聚糖酶活性是4.68IU/mL,纤维素酶活只有0.09IU/mL。     

利用啤酒废水培养生物絮凝剂产生菌的研究

从空气和活性污泥中筛选出絮凝剂产生菌,对其进行筛选并纯化得到絮凝活性较高且稳定的菌株M3。以啤酒废水为廉价培养基,对絮凝剂产生菌M3进行培养,考察外加碳源、氮源、培养基pH值、培养时间等因素对絮凝剂产生菌絮凝效果的影响。得出了M3的最佳培养条件:直接利用啤酒废水,无需另外添加碳源和氮源,只需添加0.5%的KH2PO4,温度为30℃,培养基初始pH值为8.5,培养时间为48 h,摇床转速为160 r/min。在此条件下所产生的絮凝剂对高岭土悬液絮凝率高达93.5%。     

白腐菌液体培养产生漆酶的研究

研究了碳源、氮源和综合因素对白腐菌P.conchatus液体培养产漆酶和生长情况的影响。碳源为葡萄糖,氮源为酒石酸铵时,菌株的产漆酶能力较强;麦麸为碳源,酵母膏为氮源,对菌株的生长最为有利。通过正交实验确定了最佳培养基组成为葡萄糖10 g/L,酒石酸铵5 g/L,大量元素为基础培养基的3倍,微量元素为20 mL。葡萄糖是影响菌株生长的主要因素。     

Screening and Evaluation of Polyhydroxybutyrate-Producing Strains from Indigenous Isolate Cupriavidus taiwanensis Strains

Polyhydroxyalkanoate (PHA) is a biodegradable material with many potential biomedical applications, including medical implants and drug delivery. This study developed a system for screening production strains in order to optimize PHA production in Cupriavidus taiwanensis 184, 185, 186, 187, 204, 208, 209 and Pseudomona oleovorans ATCC 29347. In this study, Sudan black B staining, Infrared (IR) and Gas Chromatography (GC) analysis indicated that the best strain for PHA synthesis is C. taiwanensis 184, which obtains polyhydroxybutyrate (PHB). Cultivation of C. taiwanensis 184 under a pH of 7.0, at 30 °C, and at an agitation rate of 200 rpm, obtained a PHB content of 10% and PHB production of 0.14 g/L. The carbon and nitrogen types selected for analysis of PHB production by C. taiwanensis 184 were gluconic acid and NH(4)Cl, respectively. Optimal carbon/nitrogen ratio for PHB production was also determined. This study demonstrated a PHB content of 58.81% and a PHB production of 2.44 g/L when the carbon/nitrogen ratio of 8/1 was selected for C. taiwanensis 184. A two-stage fermentation strategy significantly enhanced PHB content and PHB production. Under a two-stage fermentation strategy with nutrient-limited conditions, C. taiwanensis 184 obtained a PHB content of 72% and a PHB concentration of 7 g/L. Finally, experimental results confirmed that optimizing the growth medium and fermentation conditions for cultivating the indigenous C. taiwanensis 184 strain substantially elevated PHB content from 10% to 72% and PHB production from 0.14 g/L to 7 g/L, respectively.     

工业糖化酶固态发酵木薯渣制取单细胞蛋白饲料的研究

木薯是广西的主要经济作物之一,主要用于生产淀粉,其副产物木薯渣中仍残留大量淀粉,弃之会造成浪费和环境污染.本研究旨在探索一条利用木薯渣的新途径.以木薯渣为原料,利用工业糖化酶和产朊假丝酵母(Candida utilis)固态发酵,制取单细胞蛋白饲料.结果表明:通过高压灭菌预处理物料效果较好;氮源的添加量分别为:尿素3%,硫酸铵6%;通过单因素正交实验得最佳工艺条件为:糖化酶用量为4%,含水量为70%,发酵时间72 h,酵母接种量10%.粗蛋白质含量由底物的11.59%提高到产物的28.28%,提高了16.69个百分点.与其它方法比较,本方法利用工业糖化酶,发酵过程易于控制,不需要选育淀粉分解菌,使培养成本降低,具有一定的工业应用潜力.     

Solid-state production of esterase using fish processing wastes by Bacillus altitudinis AP-MSU

The bacterium Bacillus altitudinis AP-MSU, able to produce esterase was isolated from the gut of marine fish Sardinella longiceps. The esterase production was investigated in solid-state fermentation experiment using various fish processing waste meal. Among the tested fish processing wastes, red grouper waste emerged as the best source for higher esterase production. The suitable surfactant and triglyceride identified to increase the lipase production was neem oil. Effect of individual carbon and nitrogen sources supplementation on esterase production revealed that fructose and peptone aided the higher esterase production than the other tested carbon and nitrogen sources. The suitable concentration of sodium chloride for higher esterase production was at 5%. Effect of surfactants and trace elements on esterase production showed that Tween 20 and zinc sulphate, respectively produced maximum amount of esterase. The effect of physical parameters on lipase production revealed that 50 °C temperature and pH 7–8 were optimum for higher esterase production. Statistical optimization with Plackett–Burman design showed that neem oil, NaCl and fructose were found to be the most predictive factors for esterase production by this strain.     

pH值反馈控制赖氨酸补料发酵中的碳氮源补加方法

建立了一种基于发酵液pH信号反馈控制赖氨酸发酵中葡萄糖、氨和硫酸铵补加的方法,通过在不同碳源和氮源浓度下进行分批补料发酵,得到葡萄糖、氨和硫酸铵的质量消耗比为15.7:1:1.64. 按此比例配制三者的混合溶液,在补料开始后代替氨水调节pH,可在发酵过程中补加碳源和氮源. 结果表明,利用该补料方式可将葡萄糖浓度维持在8~16 g/L,铵离子浓度维持在1.52~3.38 g/L,可使赖氨酸最大浓度分别比恒基质浓度补料方式和间歇补料方式提高3.6%和17.2%,产酸率提高9.5%和28.8%,糖酸转化率提高4.9%和18.6%.     

利用农业废弃物和诱导剂提高海洋内生拟盘多毛孢菌J63的漆酶产量(英文)

Pestalotiopsis sp. J63, producing a high activity of laccase, is a new marine-derived fungus isolated from the oceanic sediment of the East China Sea. Since the marine environment is oligotrophic nutrient, marine de-rived fungi may use small amount of nutrients to grow and produce laccases. Agricultural residues that are mainly composed of lignin, cellulose and hemicellulose are difficult to be degraded and few microbes can take them as sub-strates, so they are considered as oligotrophic nutrient and have the potential to be used to produce value added products. In this study, the ability of Pestalotiopsis sp. J63 to use agricultural residues to produce laccases was tested in the submerged fermentation. The combination of 3 g·L-1 maltose and 20 g·L-1 rice straw was the best car-bon sources and 8 g·L-1 ammonium sulfate was the best nitrogen source under the condition without inducers. The effects of five inducers, the feeding time and concentration of inducer on laccase production were investigated. Adding 0.09 mmol·L-1 phenol after 24 h of incubation led to high laccase activity （5089 U·L-1）, while with 0.09 mmol·L-1 phenol in the medium and wheat bran as the nitrogen source, the laccase activity could reach 5791.7 U·L-1. Native-PAGE results showed that two laccase isozymes were present in the cultures. One existed in both in-duced and non-induced culture filtrates, while the other was only found in the fermentation with the addition of phenol, guaiacol and veratryl alcohol.     

以紫茎泽兰沼渣制作腐殖酸肥料的实验研究

以制作农残降解剂后紫茎泽兰的残渣残叶为原料,通过对其进行不同TS浓度的发酵实验,探究其适用于发酵的最佳浓度;并对发酵后沼渣的氮、磷、钾、腐殖酸等成分进行测定。结果表明,TS浓度为11.25%时产气较多;残渣腐殖酸含量与市场腐殖酸复合肥相近,但氮、钾元素相比含量较少。该实验可以为资源化利用紫茎泽兰、制作新型腐殖酸肥料提供参考依据。     

Media Optimization for Poly(β-hydroxybutyrate) Production Using Azotobacter Beijerinckii

In the present study, efforts were made to optimize the growth of Azotobacter beijerinckii in the media with essential nutrients for production of PHB. The effects of temperature, agitation rate, carbon and nitrogen source concentrations on microbial grows and PHB production were investigated. Glucose and ammonium chloride were used as carbon and nitrogen source, respectively. The optimal temperature for the growth and PHB synthesis appeared to be 30°C. However, over the range of 25–38°C, the effect of temperature was not very significant. Fermentation under controlled conditions such as agitation rate of 250 rpm, incubated inoculums for 15 h, fermentation temperature at 30°C, and fermentation time of 72 h was performed. Glucose concentration of 30 to 60 g/l with increment of 10 g/l and ammonium chloride in the range of 0.5 to 2 g/l with increment of 0.5 g/l were utilized as carbon and nitrogen sources, respectively. The PHB production was maximized at the desired concentration of glucose (50 g/l) and ammonium chloride (1.5) for carbon and nitrogen sources, respectively. A. beijerinckii exhibited maximum biomass of 17.31 g/l with a PHB concentration of 5.84 g/l. Under the best conditions, PHB constituted up to 34% of dry cell mass after 64 h of culture. The average biomass yield coefficient on glucose was about 0.57 kg/kg.     

Improvement of fermentative production of exopolysaccharides from Aureobasidium pullulans under various conditions

The optimization of exopolysaccharide (EPS) production was investigated in the polymorphic fungal strain of Aureobasidium pullulans (KCTC 6081) with varying pH, nutrients concentration, and mixing parameters in batch fermentation condition. The maximum production of EPS (～7.5 g/L) was observed at pH 4, while optimum nutrient concentration of carbon (sucrose), nitrogen (NaNO₃), phosphorous (K₂HPO₄), and ascorbic acid was 50 g/L, 5 g/L, 1 g/L and 2 g/L, respectively. Interestingly, EPS productivity under non pH controlled fermentation conditions was 0.12 g/L/h with 400 rpm mixing, while under a controlled pH of 4, the EPS productivity was 0.21 g/L/h with 600 rpm, respectively. The fed-batch fermentation increased the EPS productivity up to 0.345 g/L/h with changing mixing conditions from 200 to 600 rpm and reached 47 g/L with 88% pullulan. Thus, pH and mixing were the key parameters for enhancing EPS production from A. pullulans. It is expected that these optimized parameters can be well used for enhanced industrial production of pullulan.     

酿酒酵母工程菌发酵产青蒿酸的工艺优化

通过酿酒酵母工程菌(Saccharomyces cerevisiae 1211)发酵制备青蒿酸,并通过单因素实验和响应面优化,考察了发酵温度、pH、半乳糖质量浓度、发酵碳源及发酵氮源等对青蒿酸发酵产量的影响。结果表明:在发酵温度30℃,发酵培养基初始pH=5.5,发酵培养基中蔗糖质量浓度91.8 g/L,半乳糖质量浓度10.1 g/L,硫酸铵质量浓度10.3 g/L,磷酸二氢钾质量浓度8.7 g/L的条件下,青蒿酸发酵产量可达(1529.7±12.6)mg/L,与未优化时的发酵产量相比,提升了67.1%。