Effects of Octadecanoylsucrose Derivatives on the Production of Inulinase by Apergillus niger SL-09

Summary The effect of the addition of octadecanoylsucrose esters to the growth medium on the production of inulinase by Aspergillus niger SL-09 was studied in batch culture using shake flasks. The activities of inulinase in vitro and in vivo formed by Aspergillus niger SL-09 was enhanced dramatically by the addition of sucrose ester S-770 to the medium, and it was confirmed that sucrose ester acted as a very efficient inducer for inulinase production. As a result, with the addition of 6 g sucrose ester l⁻¹ at the beginning of the culture, the enzyme activities were enhanced near 7-fold higher than that obtained in the basal medium.     

响应面方法优化菊粉酶液体发酵培养基的研究

目的：为提高菊粉酶的产量。方法：运用Plackett-Burman设计法和响应面分析法,对Kluyveromyces S120液体发酵生产菊粉酶的培养基进行了优化。首先通过Plackett-Burman方法对8个相关影响因素的效应进行评价,并筛选出了有显著正效应的菊芋粉、玉米浆、(NH_4)H_2PO_4等三个因素,其他五个因素没有显著影响。然后根据Box-Behnken的中心组合设计实验和响应面分析方法确定了上述三个主要影响因素的最佳浓度。结果：在优化培养基下,菊粉酶产量为102．82u/mL,是优化前的2．1倍。     

A New Method for the Preparation of Minicells for Physiological Studies

A procedure has been developed for preparing minicells that does not rely on sucrose gradients in a rate-zone centrifuge. In the presence of low levels (10 units/ml) of penicillin, the contaminating bacteria present in minicell cultures after low-speed differential centrifugation are turned into long filamentous cells and can be killed by sonic treatment. An additional low-speed centrifugation (2, 000 g for 5 min) yields purified minicells with less than one contaminating cell per 10 minicells.     

复合诱变在青霉高产菊粉酶菌株选育中的应用

以Penicillium sp．B01为出发菌株,经吖黄素或DES（硫酸二乙酯）分别与^60Co-γ射线对其孢子悬液进行复合诱变。经过初筛和复筛,在30μg／mL吖黄素诱变时间2h,剂量率为4．11Gy／min的^60Co-γ射线辐射使累计剂量为20．55Gy复合诱变的条件下,筛选出一株菊粉酶活比出发菌株高32％的突变菌株B01-A13-Co31。经同工酶电泳验证,变异株与出发菌株相比酶带有所变化。将此菌种连续传代6次进行产酶性能的稳定性测定,表明此菌株具有良好的遗传稳定性。     

黑曲霉产菊粉酶菌株的选育及培养基优化

为获得高产菊粉酶的黑曲霉菌株,以Aspergillus niger YH-1为出发菌株,经过亚硝基胍(NTG)诱变,以高温高菊芋粉相结合的方式进行梯度驯化,选育出一株产菊粉酶菌株YH-3,并运用响应面实验方法对该菌株的培养基进行优化。确定了最佳培养基组成:菊芋粉25.2 g/L、豆饼粉40 g/L、蔗糖酯4.9 g/L、NaCl 5.5 g/L。发现内切菊粉酶活力(I)由60.9 U/mL提高到165.0 U/mL,比出发菌株提高了1.7倍。研究证明蔗糖酯对于黑曲霉YH-3发酵产菊粉酶是一种有效的促进剂。     

云南省部分地区产菊粉酶霉菌的初步调查

从昆明,石林,西双版纳,思茅等地区的菊科植物根系上群,昆明,石林地区市售洋姜表土,洋姜提取液中污染菌及洋姜表面霉烂处,分离纯化产菊粉酶菌株,共获得105株霉菌,对以上菌株进行了初步鉴定,对产酶情况进行了初步的研究。发现分属于拟青霉属（Paecilomyces sp.),头孢霉属（Cephalosporium sp.),阜孢霉属（Papularia sp.),丝衣霉属（Byssochlamys sp.),指轮枝孢霉属（Stachylidium sp.)。刚毛菌属（Lacellina sp.)交链孢属（Alternaria sp.),地霉属（Geotrichun sp.)毛霉属（Mucor sp.),枝孢霉属（Claddosporium sp.),金孢霉属（Chrysosporium sp.),镰刀菌属（Fusarium sp.),根霉属（Rhizopus sp．）等13个避的霉菌,是国内未见报道的新的菊粉酶产生菌。     

1株产菊粉酶菌株的筛选、鉴定及发酵条件优化

筛选分离可以分解菊芋中菊糖的菌株。采用平板稀释法从土样中筛选出能够分解菊芋中菊糖的菌株,并从中得到1株酶活较高的真菌A-15,经菌落观察及采用18S rRNA基因测序鉴定,研究不同因素对菌株菊粉酶活力的影响。通过对分离得到的菌株形态观察及分子鉴定后,确定菌株A-15为黑曲霉(Aspergillus niger)。通过正交试验优化菌株A-15的发酵条件分析结果显示,菌株A-15产菊粉酶最优条件:最佳氮源为酵母膏,氮源量1.0%,氯化钠0.5%,磷酸氢二钾0.3%,菊芋汁定容,初始pH 6,培养温度30℃,摇瓶发酵6 d。结果表明菌株A-15具有很好地降解菊芋中菊糖的性能。     

Ethanol fermentation with Kluyveromyces marxianus from Jerusalem artichoke grown in salina and irrigated with a mixture of seawater and freshwater

Aims: To study fuel ethanol fermentation with Kluyveromyces marxianus ATCC8554 from Jerusalem artichoke (Helianthus tuberosus) grown in salina and irrigated with a mixture of seawater and freshwater. Methods and Results: The growth and ethanol fermentation of K. marxianus ATCC8554 were studied using inulin as substrate. The activity of inulinase, which attributes to the hydrolysis of inulin, the main carbohydrate in Jerusalem artichoke, was monitored. The optimum temperatures were 38°C for growth and inulinase production, and 35°C for ethanol fermentation. Aeration was not necessary for ethanol fermentation with the K. marxianus from inulin. Then, the fresh Jerusalem artichoke tubers grown in salina and irrigated with 25% and 50% seawater were further examined for ethanol fermentation with the K. marxianus, and a higher ethanol yield was achieved for the Jerusalem artichoke tuber irrigated with 25% seawater. Furthermore, the dry meal of the Jerusalem artichoke tubers irrigated with 25% seawater was examined for ethanol fermentation at three solid concentrations of 200, 225 and 250 g l^?1, and the highest ethanol yield of 0·467, or 91·5% of the theoretical value of 0·511, was achieved for the slurry with a solid concentration of 200 g l^?1. Conclusions: Halophilic Jerusalem artichoke can be used for fuel ethanol production. Significance and Impact of the Study: Halophilic Jerusalem artichoke, not competing with grain crops for arable land, is a sustainable feedstock for fuel ethanol production.     

Studies on enzymic methods for extraction of inulin from Jerusalem artichokes

The release of inulin, d-fructose and protein from Jerusalem artichokes has been studied under diffusion and maceration conditions. The effects on release of added inulinase (2,1-β-d-fructan fructanohydrolase, EC 3.2.1.7), protease and cellulase [see 1,4-(1,3;1,4)-β-d-glucan 4-glucanohydrolase, EC 3.2.1.4] enzymes have been investigated. The results indicate that added enzymes do not improve the efficiency of inulin or d-fructose release and that mechanical methods represent the most efficient means of carbohydrate solubilization. Treatment of plant extracts with inulinase is shown to have the disadvantage of increasing peptide solubilization. The potential for improved inulin solubilization by use of endo-inulinases is discussed.     

Improvement of inulinase stability of calcium alginate immobilized Kluyveromyces marxianus cells by treatment with hardening agents

To improve the inulinase (2,1-β-d-fructan fructanohydrolase, EC 3.2.1.7) stability of calcium alginate-immobilized Kluyveromyces marxianus cells, treatment with hardening agents has been investigated. Treatment of immobilized cells with some polycationic polymers resulted in little decrease in volumetric reactor productivity, but was most effective in increasing the inulinase stability of the immobilized cells. Inulinase stability of glutaraldehyde-hardened immobilized cells increased two-fold, and for hexamethylenediamine + glutaraldehyde and polyethyleneimine + glutaraldehyde-hardened cells increased six-fold compared with that of the unhardened cells.