碱性β-聚糖酶产生菌选育及产酶条件优化

从碱性土样中分离到数十株产碱性β聚糖酶类的细菌,经摇瓶反复筛选后,得到一株碱性β聚糖酶产量较高的耐碱性细菌．经初步鉴定,属短小芽孢杆菌（Ｂａｃｉｌｕｓｐｕｍｉｌｕｓ）．其纤维素酶作用的最适ｐＨ为７．６,最适θ为６０℃,木聚糖酶的作用最适ｐＨ为９．０,最适θ为５５℃．该菌的最适生长ｐＨ为８．０,最适产酶θ为２８～３２℃．木聚糖与山梨糖分别是木聚糖酶和纤维素酶的良好诱导物．以麸皮为碳源,产酶的最适浓度为５％．添加尿素和（ＮＨ４）２ＳＯ４为氮源可提高纤维素酶酶活２倍,木聚糖酶酶活１倍．发酵周期为６０ｈ,纤维素酶酶活最高可达１．２１ＩＵｍＬ－１,木聚糖酶酶活可达４３ＩＵｍＬ－１．     

苏云金芽孢杆菌生物杀虫剂发酵生产的影响因素及其工艺选择

化学杀虫剂的长期使用给生态环境造成了严重破坏,也使害虫种群的抗药性日益提高,生物杀虫剂以其＂绿色环保＂的特点引起人们的广泛关注。其中,苏云金芽孢杆菌（Bacillus thuringiensis）制剂是目前世界上产量最大、应用最广的生物杀虫剂。它对鳞翅目、双翅目、鞘翅目、螨类等许多有害昆虫有毒杀作用,而对人类、动物和农作物无害。长期以来,人们一直致力于苏云金芽孢杆菌发酵过程的研究,以期获得高毒效的生物杀虫剂产品。本文首先对苏云金芽孢杆菌发酵生产的各种影响因素进行了综合分析,将影响因素分为培养条件和培养基组分两类,得出最佳培养条件为温度：（30±1）℃,pH：7.0±0.1,搅拌速度：400～600r·min-1,通气量：1∶0.6～1.2（发酵培养基体积与每分钟通入空气的体积之比）,接种时间：对数期初;最佳培养基配比为碳氮比：8～10∶1,无机盐含量：KH2PO4或K2HPO4为0.075%～0.2%;MgSO4·7H2O为0.075%～0.3%;CaCO3为0.075%～0.15%;MnSO4·H2O、FeSO4·7H2O各为0.002%。其次,对当前研究与工业化生产中的各种发酵工艺进行了评述,总结了现有发酵工艺的优缺点。在现有研究基础上,降低培养基原料成本、改进发酵工艺和采用基因学手段构建高效工程菌株将成为未来研究热点。     

红树林沉积物Bt菌株的分离与cry基因型的鉴定（Isolation of Bacillus thuringiensis Strains from Mangrove Sediment Samples and Identification for cry Genes）

从收集自泉州洛江红树林保护区的400个沉积物土壤样品中,分离到18株苏云金芽孢杆菌(Bacillus thuringiensis,Bt).利用PCR-RFLP体系对Bt菌株杀虫晶体蛋白的基因型进行了鉴定,发现15株Bt含有cry1基因,4株含有cry2基因,有3株分别含有cry7、cry8、cry9基因.克隆了一个新型的cry2Ab基因,其编码蛋白与现有Cry2Ab型杀虫晶体蛋白的最高同源性为95%.     

一株芽孢杆菌产生物絮凝剂去除赤潮微藻的研究

以赤潮微藻米氏凯伦藻(Karenia mikimotoi)和塔玛亚历山大藻(Alexandrium tamarense)为受试对象,对一株芽孢杆菌WZ01所产的生物絮凝剂去除赤潮微藻的性能进行了研究。结果表明:芽孢杆菌WZ01所产生物絮凝剂对人工培养的米氏凯伦藻和塔玛亚历山大藻藻液均有去除能力,适宜的絮凝剂质量浓度为40.0~60.0 mg/L,絮凝时间为90~120 min,pH=7.5~8.5,在20~40℃范围内生物絮凝剂对2种微藻的絮凝率无显著区别;Ca~(2+)和Mg~(2+)浓度对生物絮凝剂去除2种微藻有一定影响,适宜浓度为4.0~6.0 mmol/L;适宜条件下生物絮凝剂对人工培养的米氏凯伦藻和塔玛亚历山大藻藻液的絮凝率均超过80.0%。研究表明,芽孢杆菌WZ01所产的生物絮凝剂在去除赤潮微藻方面有应用潜力。     

产木聚糖酶嗜碱细菌的筛选及产酶条件研究

利用透明圈法筛选到一株产木聚糖酶的嗜碱芽孢杆菌NT 9,该菌株产木聚糖酶为组成型。正交设计实验结果表明 ,合适的产酶条件为 :木糖 15 .0g/L ,硫酸铵 2 .5 g/L ,Tween 80 2 0 g/L ,K2 HPO4 1.0g/L ,MgSO4 ·7H2 O 0 .2g/L ,pH值 10 .0 ,3 7℃ ,2 0 0r/min ,振荡培养 72h。其木聚糖酶活力可达到 6.3 6IU/mL。     

好氧砷还原菌对吸附态砷迁移转化的影响

利用静态和动态土柱实验初步研究了好氧砷还原菌Bacillus sp. SXB在砷迁移转化过程中的作用。结果表明,在静态实验中,菌株SXB可以有效还原吸附在针铁矿上的五价砷;在动态土柱实验中,由于原土中砷是以三价砷形式存在,因此,不论是空白对照还是菌株SXB加入处理,出水中砷均为三价砷;当孔隙体积大于100时,铁的溶出开始增加。菌株SXB的加入,轻微地促进了土壤中砷的释放,而对铁的释放并没有明显影响。     

一株鱼粉加工硫化氢恶臭气体脱除菌株的分离与鉴定

鱼粉是水产动物、及畜禽动物养殖饲料中无法替代的优质动物蛋白源。然而因加工原料新鲜度不断下降、腐败等原因,鱼粉加工过程中会释放出大量的硫化氢恶臭气体,导致周边环境的严重污染。取鱼粉厂污水沉淀池中的污泥,对脱硫微生物进行富集、分离、纯化、检测脱硫效率,得到一株高效脱硫菌株XJ-2,其脱硫效率达87.1%。通过菌株形态观察、生理生化实验、API 50CHB试剂条检测及16S rDNA测序鉴定,结果确定该菌为蜡状芽孢杆菌(Bacillus cereus)。研究菌株XJ-2生长特性,发现菌株对环境中的pH值耐受力较强,不耐高温,正好避开鱼粉厂休业期,且经生长曲线研究发现该菌的稳定期较长,具有良好的应用前景。此研究将为生物法脱除鱼粉加工恶臭气体的应用提供一定的理论基础。     

微波无极紫外方法对二沉池出水的消毒研究

采用自制微波无极紫外连续式消毒反应器,对二沉池出水进行消毒.紫外光源为自制圆柱形无极灯,灯内充汞10mg、氩气压强666.61Pa紫外光强5.07mW/cm²,进水流量0.072L/s,出水细菌总数2CFU/mL,大肠菌群数0.微波无极紫外作用过程中,微波破坏细胞壁细胞膜结构使蛋白质、K⁺等胞内物质产生渗漏,这种不可逆的破坏过程有效抑制紫外光复活现象.     

Controlling bacterial leaf blight of rice and enhancing the plant growth with endophytic and rhizobacterial Bacillus strains

This study was conducted to assess efficacy of biological control against bacterial leaf blight (BLB) of rice produced by Xanthomonas oryzae pv. oryzae. Five endophytic strains (A1, A2, A3, A13 and A15) and two rhizospherial Bacilli (D29 and H8) were tested for their antagonistic activities against BLB in vitro and in vivo. All seven strains showed high potential of antagonistic activity against X. oryzae pv. oryzae and three phytopathogenic fungi in vitro. Test of 16SrRNA gene sequence were assigned isolates A1, A3 and A13 as Bacillus amyloliquefaciens while isolates A2 and A15 as B. methylotrophicus and B. subtilis, respectively. In greenhouse, four strains of displayed 50.29%–57.86% inhibition rate against the pathogen and significantly increased plant fresh weight from 50.03% to 73.11% and dry weight from 64.11% to 86.65% in treated rice plants. In addition, these strains demonstrated strong capability to produce indole-3-acetic acid, siderophores, solubilizing phosphate and also colonize roots. Real-time quantitative polymerase chain reaction revealed that expression of defense-related genes including OsAOS2, OsJMT1, OsNPR1 and OsPR1b were significantly up-regulated in leaves of D29-exposed rice plants, suggesting that treatment of rice with D29 suppressed BLB through systemic activation of the plant defense system. Therefore, data suggest that Bacillus isolates A13, A15, D29 and H8 support effective antagonistic activity against BLB under greenhouse conditions in addition to their potential to promote growth of rice plants.     

Assessment of the in vivo genotoxicity of a new formulation of Bacillus thuringiensis var israelensis SH-14

Biological control agents have become a useful alternative for the reduction of the use of chemical insecticides. LABIOFAM (Cuba) is developing a new formulation of a biolarvicide that possesses as active biological agent Bacillus thuringiensis var israelensis serotype H14. In order to evaluate the genotoxicity of this new formulation, an in vivo battery test was used: micronucleus (MN), chromosome aberrations (CAs), and sperm morphology (SM) assays. A dose of 6.45?×?10⁸ spores was administered per animal via oral administration. Bone marrow cells were collected 24?h after a two day treatment for the MN assay, and 24?h after a unique treatment for the CA assay, using cyclophosphamide as the positive control. Sperm cells were collected at 5 weeks from the first of five administrations for the SM test, using acrylamide as positive control. Bacillus thuringiensis var israelensis serotype H14 failed to show either a significative increase of micronucleated polychromatic erythrocytes, chromosomal aberrations, or sperm abnormalities. Acute oral administration of a high dose of Bacillus thuringiensis var israelensis serotype H14 did not produce mutagenic effects in bone marrow or sperm cells.