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Optimization of Medium Composition for the Production of Neomycin by Streptomyces fradiae NCIM 2418 in Solid State Fermentation. 传统链霉菌NCIM 2418固态发酵生产新霉素的培养基组成优化。
Pub Date : 2014-01-01 Epub Date: 2014-06-09 DOI: 10.1155/2014/674286
B M Vastrad, S E Neelagund

Neomycin production of Streptomyces fradiae NCIM 2418 was optimized by using response surface methodology (RSM), which is powerful mathematical approach comprehensively applied in the optimization of solid state fermentation processes. In the first step of optimization, with Placket-Burman design, ammonium chloride, sodium nitrate, L-histidine, and ammonium nitrate were established to be the crucial nutritional factors affecting neomycin production significantly. In the second step, a 2(4) full factorial central composite design and RSM were applied to determine the optimal concentration of significant variable. A second-order polynomial was determined by the multiple regression analysis of the experimental data. The optimum values for the important nutrients for the maximum were obtained as follows: ammonium chloride 2.00%, sodium nitrate 1.50%, L-histidine 0.250%, and ammonium nitrate 0.250% with a predicted value of maximum neomycin production of 20,000 g kg(-1) dry coconut oil cake. Under the optimal condition, the practical neomycin production was 19,642 g kg(-1) dry coconut oil cake. The determination coefficient (R (2)) was 0.9232, which ensures an acceptable admissibility of the model.

采用响应面法(RSM)对传统链霉菌NCIM 2418生产新霉素进行了优化。响应面法是一种综合应用于固态发酵工艺优化的强大数学方法。在第一步优化中,采用Placket-Burman设计,确定了氯化铵、硝酸钠、l -组氨酸和硝酸铵是影响新霉素产量的关键营养因子。第二步,采用2(4)全因子中心组合设计和RSM来确定显著变量的最佳浓度。通过对实验数据的多元回归分析,确定了一个二阶多项式。得到的重要营养物质的最适用量为:氯化铵2.00%、硝酸钠1.50%、l -组氨酸0.250%、硝酸铵0.250%,预计最大新霉素产量为2万g kg(-1)干椰子油饼。在此条件下,实际新霉素产量为19642 g kg(-1)干椰子油饼。决定系数(R(2))为0.9232,保证了模型的可接受性。
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引用次数: 14
Solid State Fermentation of a Raw Starch Digesting Alkaline Alpha-Amylase from Bacillus licheniformis RT7PE1 and Its Characteristics. 地衣芽孢杆菌RT7PE1消化碱性α -淀粉酶的生淀粉固态发酵及其特性研究
Pub Date : 2014-01-01 Epub Date: 2014-01-21 DOI: 10.1155/2014/495384
Romana Tabassum, Shazia Khaliq, Muhammad Ibrahim Rajoka, Foster Agblevor

The thermodynamic and kinetic properties of solids state raw starch digesting alpha amylase from newly isolated Bacillus licheniformis RT7PE1 strain were studied. The kinetic values Q p , Y p/s , Y p/X , and q p were proved to be best with 15% wheat bran. The molecular weight of purified enzyme was 112 kDa. The apparent K m and V max values for starch were 3.4 mg mL(-1) and 19.5 IU mg(-1) protein, respectively. The optimum temperature and pH for α -amylase were 55°C, 9.8. The half-life of enzyme at 95°C was 17h. The activation and denaturation activation energies were 45.2 and 41.2 kJ mol(-1), respectively. Both enthalpies (ΔH (∗)) and entropies of activation (ΔS (∗)) for denaturation of α -amylase were lower than those reported for other thermostable α -amylases.

研究了新分离的地衣芽孢杆菌RT7PE1菌株的固态原料淀粉消化α淀粉酶的热力学和动力学性质。在15%麦麸条件下,qp、yp /s、yp /X和qp的动力学值最好。纯化酶分子量为112 kDa。淀粉的表观K - m和V - max分别为3.4 mg mL(-1)和19.5 IU mg(-1)蛋白。α -淀粉酶最适温度为55℃,pH为9.8℃。酶在95℃时的半衰期为17h。活化活化能和变性活化能分别为45.2和41.2 kJ mol(-1)。α -淀粉酶变性的焓(ΔH(∗))和激活熵(ΔS(∗))均低于其他耐热性α -淀粉酶。
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引用次数: 14
Isolation and characterization of bacteriophages infecting nocardioforms in wastewater treatment plant. 污水处理厂诺心虫噬菌体的分离与鉴定。
Pub Date : 2014-01-01 Epub Date: 2014-07-22 DOI: 10.1155/2014/151952
Krishna Khairnar, Preeti Pal, Rajshree H Chandekar, Waman N Paunikar

Activated sludge plants (ASP) are associated with the stable foaming problem worldwide. Apart from the physical and chemical treatment methods, biological treatment method has been least explored and may prove to be a novel and ecofriendly approach to tackle the problem of stable foam formation. In ASP Nocardia species are commonly found and are one of the major causes for forming sticky and stable foam. This study describes the isolation and characterization of three Nocardia bacteriophages NOC1, NOC2, and NOC3 for the control of Nocardia species. The bacteriophages isolated in this study have shown promising results in controlling foam producing bacterial growth under laboratory conditions, suggesting that it may prove useful in the field as an alternative biocontrol agent to reduce the foaming problem. To the best of our knowledge to date no work has been published from India related to biological approach for the control of foaming.

活性污泥厂(ASP)在世界范围内与稳定发泡问题密切相关。除物理和化学处理方法外,生物处理方法探索最少,可能被证明是解决稳定泡沫形成问题的一种新颖而环保的方法。在ASP中,诺卡菌种类是常见的,是形成粘性和稳定泡沫的主要原因之一。本研究对诺卡菌的三种噬菌体NOC1、NOC2和NOC3进行了分离和鉴定。本研究中分离的噬菌体在控制实验室条件下产生泡沫的细菌生长方面显示出有希望的结果,这表明它可能在现场被证明是一种可替代的生物防治剂,以减少泡沫问题。据我们所知,迄今为止,印度还没有发表过与控制泡沫的生物方法有关的工作。
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引用次数: 21
Parametric Optimization of Cultural Conditions for Carboxymethyl Cellulase Production Using Pretreated Rice Straw by Bacillus sp. 313SI under Stationary and Shaking Conditions. 静止和振荡条件下芽孢杆菌313SI预处理稻草生产羧甲基纤维素酶培养条件的参数优化
Pub Date : 2014-01-01 Epub Date: 2014-04-29 DOI: 10.1155/2014/651839
Varsha Goyal, Arpana Mittal, Anish Kumari Bhuwal, Gulab Singh, Anita Yadav, Neeraj Kumar Aggarwal

Carboxymethyl cellulase (CMCase) provides a key opportunity for achieving tremendous benefits of utilizing rice straw as cellulosic biomass. Out of total 80 microbial isolates from different ecological niches one bacterial strain, identified as Bacillus sp. 313SI, was selected for CMCase production under stationary as well as shaking conditions of growth. During two-stage pretreatment, rice straw was first treated with 0.5 M KOH to remove lignin followed by treatment with 0.1 N H2SO4 for removal of hemicellulose. The maximum carboxymethyl cellulase activity of 3.08 U/mL was obtained using 1% (w/v) pretreated rice straw with 1% (v/v) inoculum, pH 8.0 at 35°C after 60 h of growth under stationary conditions, while the same was obtained as 4.15 U/mL using 0.75% (w/v) pretreated substrate with 0.4% (v/v) inoculum, pH 8.0 at 30°C, under shaking conditions of growth for 48 h. For maximum titre of CMCase carboxymethyl cellulose was optimized as the best carbon source under both cultural conditions while ammonium sulphate and ammonium nitrate were optimized as the best nitrogen sources under stationary and shaking conditions, respectively. The present study provides the useful data about the optimized conditions for CMCase production by Bacillus sp. 313SI from pretreated rice straw.

羧甲基纤维素酶(CMCase)为实现利用稻草作为纤维素生物质的巨大效益提供了关键机会。在来自不同生态位的80株分离菌中,选择了一株被鉴定为芽孢杆菌sp. 313SI的菌株,在静止和振动生长条件下生产CMCase。在两阶段预处理中,先用0.5 M KOH处理秸秆脱除木质素,再用0.1 N H2SO4处理秸秆脱除半纤维素。采用1% (w/v)预处理稻秆,接种量为1% (v/v), pH为8.0,固定条件下生长60 h,羧甲基纤维素酶活性最高为3.08 U/mL;采用0.75% (w/v)预处理底物,接种量为0.4% (v/v), pH为8.0,接种量为30℃,振荡条件下生长48 h,羧甲基纤维素酶活性最高为4.15 U/mL。在两种培养条件下,以羧甲基纤维素为最佳碳源,以硫酸铵和硝酸铵为最佳氮源。本研究为芽孢杆菌313SI利用预处理稻草生产CMCase的优化条件提供了有益的数据。
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引用次数: 30
Overcome of Carbon Catabolite Repression of Bioinsecticides Production by Sporeless Bacillus thuringiensis through Adequate Fermentation Technology. 利用适当的发酵技术克服苏云金芽孢杆菌碳分解代谢抑制生物杀虫剂生产。
Pub Date : 2014-01-01 Epub Date: 2014-09-17 DOI: 10.1155/2014/698587
Saoussen Ben Khedher, Samir Jaoua, Nabil Zouari

The overcoming of catabolite repression, in bioinsecticides production by sporeless Bacillus thuringiensis strain S22 was investigated into fully controlled 3 L fermenter, using glucose based medium. When applying adequate oxygen profile throughout the fermentation period (75% oxygen saturation), it was possible to partially overcome the catabolite repression, normally occurring at high initial glucose concentrations (30 and 40 g/L glucose). Moreover, toxin production yield by sporeless strain S22 was markedly improved by the adoption of the fed-batch intermittent cultures technology. With 22.5 g/L glucose used into culture medium, toxin production was improved by about 36% when applying fed-batch culture compared to one batch. Consequently, the proposed fed-batch strategy was efficient for the overcome of the carbon catabolite repression. So, it was possible to overproduce insecticidal crystal proteins into highly concentrated medium.

研究了苏云金芽孢杆菌(Bacillus thuringiensis) S22菌株在3 L全控发酵罐中利用葡萄糖培养基生产生物杀虫剂时分解代谢物抑制的问题。当在整个发酵期间应用足够的氧气(75%的氧饱和度)时,可以部分克服分解代谢物抑制,通常发生在高初始葡萄糖浓度(30和40 g/L葡萄糖)下。采用间歇式间歇培养技术可显著提高无孢子菌株S22的产毒量。当培养基中添加22.5 g/L葡萄糖时,与单批培养相比,分批培养的毒素产量提高了约36%。因此,提出的补料间歇策略是有效的克服碳分解代谢抑制。因此,在高浓度培养基中过量产生杀虫晶体蛋白是可能的。
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引用次数: 10
Antidepressant Fluoxetine Modulates the In Vitro Inhibitory Activity of Buffalo Brain Cystatin: A Thermodynamic Study Using UV and Fluorescence Techniques. 抗抑郁药氟西汀调节水牛脑胱抑素体外抑制活性:紫外和荧光技术的热力学研究
Pub Date : 2014-01-01 Epub Date: 2014-07-24 DOI: 10.1155/2014/319397
Fakhra Amin, Bilqees Bano

Cystatins constitute a superfamily of homologous proteins. The major role of cystatins is to regulate the unwanted proteolysis and to protect the organism against endogenous proteases released from lysosomes, invading microorganisms and parasites that use cysteine proteases to enter the body. Imbalance in regulation of proteolytic activity may lead to a wide range of human diseases. An enormous progress has been made in understanding of protein degradation process under normal and pathological conditions; infact proteases are now clearly viewed as important drug targets. Fluoxetine a selective serotonin reuptake inhibitor (SSRI) is an antidepressant. It is used to treat major depressive disorders. In the present study binding of fluoxetine to cystatin was studied by UV and fluorescence quenching technique. Intrinsic fluorescence of fluoxetine complexed with purified buffalo brain cystatin (BC) was measured by selectively exciting the tryptophan residues. Gradual quenching was observed on complex formation. When cystatin was added to fluoxetine solutions at a molar ratio of 1 : 0.5, it not only quenched more than half of its fluorescence but also reduced the activity of cystatin. Stern-Volmer plots obtained from experiments carried out at 25(°)C showed the quenching of fluorescence to be a collisional phenomenon. Our results suggest the prime binding site for fluoxetine on BC to be at or near tryptophan residues. Fluoxetine quenched the fluorescence by a static process, which specifically indicates the formation of a complex.

半胱抑素构成一个同源蛋白超家族。半胱氨酸蛋白酶的主要作用是调节不需要的蛋白质水解,保护生物体免受溶酶体释放的内源性蛋白酶、利用半胱氨酸蛋白酶进入体内的入侵微生物和寄生虫的侵害。蛋白质水解活性调节的不平衡可能导致广泛的人类疾病。对正常和病理条件下蛋白质降解过程的认识取得了巨大进展;事实上,蛋白酶现在被清楚地视为重要的药物靶点。氟西汀是一种选择性血清素再摄取抑制剂(SSRI),是一种抗抑郁药。它被用来治疗重度抑郁症。本研究采用紫外和荧光猝灭技术研究了氟西汀与胱抑素的结合。用选择性激发色氨酸残基的方法测定了氟西汀与纯化的水牛脑胱抑素(BC)配合物的固有荧光。在络合物形成过程中观察到逐渐淬灭。将胱抑素以1:50 . 0的摩尔比加入到氟西汀溶液中,不仅淬灭了一半以上的荧光,还降低了胱抑素的活性。在25(°)C下进行的实验得到的Stern-Volmer图显示荧光猝灭是一种碰撞现象。我们的结果表明氟西汀在BC上的主要结合位点在色氨酸残基上或附近。氟西汀通过静态过程淬灭荧光,这特别表明形成了络合物。
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引用次数: 1
Cloning, Sequencing, and In Silico Analysis of β-Propeller Phytase Bacillus licheniformis Strain PB-13. 地衣芽孢杆菌菌株PB-13 β-螺旋桨植酸酶的克隆、测序及分析
Pub Date : 2014-01-01 Epub Date: 2014-04-24 DOI: 10.1155/2014/841353
Vinod Kumar, Gopal Singh, Punesh Sangwan, A K Verma, Sanjeev Agrawal

β -Propeller phytases (BPPhy) are widely distributed in nature and play a major role in phytate-phosphorus cycling. In the present study, a BPPhy gene from Bacillus licheniformis strain was expressed in E. coli with a phytase activity of 1.15 U/mL and specific activity of 0.92 U/mg proteins. The expressed enzyme represented a full length ORF "PhyPB13" of 381 amino acid residues and differs by 3 residues from the closest similar existing BPPhy sequences. The PhyPB13 sequence was characterized in silico using various bioinformatic tools to better understand structural, functional, and evolutionary aspects of BPPhy class by multiple sequence alignment and homology search, phylogenetic tree construction, variation in biochemical features, and distribution of motifs and superfamilies. In all sequences, conserved sites were observed toward their N-terminus and C-terminus. Cysteine was not present in the sequence. Overall, three major clusters were observed in phylogenetic tree with variation in biophysical characteristics. A total of 10 motifs were reported with motif "1" observed in all 44 protein sequences and might be used for diversity and expression analysis of BPPhy enzymes. This study revealed important sequence features of BPPhy and pave a way for determining catalytic mechanism and selection of phytase with desirable characteristics.

β -螺旋桨植酸酶(BPPhy)广泛存在于自然界中,在植酸-磷循环中起重要作用。本研究在大肠杆菌中表达了地衣芽孢杆菌BPPhy基因,其植酸酶活性为1.15 U/mL,比活性为0.92 U/mg蛋白。所表达的酶是全长ORF“PhyPB13”,包含381个氨基酸残基,与现有最接近的类似BPPhy序列相差3个残基。利用多种生物信息学工具,通过序列比对和同源性搜索、系统发育树构建、生化特征变化、基序和超家族分布,对PhyPB13序列进行了硅质化表征,以更好地了解BPPhy类的结构、功能和进化方面的信息。在所有序列中,在它们的n端和c端都观察到保守位点。该序列中不存在半胱氨酸。总体而言,在生物物理特征变化的系统发育树上观察到三个主要集群。44个蛋白序列中共发现10个基序,其中基序“1”可用于BPPhy酶的多样性和表达分析。该研究揭示了BPPhy的重要序列特征,为确定催化机制和选择具有理想特性的植酸酶铺平了道路。
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引用次数: 14
Characterization of β -Glucosidase Produced by Aspergillus niger under Solid-State Fermentation and Partially Purified Using MANAE-Agarose. 黑曲霉固态发酵产β -葡萄糖苷酶的特性及部分纯化
Pub Date : 2014-01-01 Epub Date: 2014-04-01 DOI: 10.1155/2014/317092
Anderson Baraldo Junior, Diogo G Borges, Paulo W Tardioli, Cristiane S Farinas

β -Glucosidase (BGL) is a hydrolytic enzyme with specificity for a wide variety of glycoside substrates, being an enzyme with a large range of biotechnological applications. However, enzyme properties can be different depending both on the microorganism and the cultivation procedure employed. Therefore, in order to explore potential biocatalytical applications of novel enzymes, their characterization is essential. In this work, a BGL synthesized by a selected strain of Aspergillus niger cultivated under solid-state fermentation (SSF) was partially purified and fully characterized in terms of optimum pH, temperature, and thermostability. The single-step purification using MANAE-agarose in a chromatographic column yielded an enzyme solution with specific activity (17.1 IU/mg protein) adequate for the characterization procedures. Electrophoresis SDS-PAGE and size-exclusion chromatography analysis resulted in an estimated molecular mass of 60 kDa. Higher enzyme activities were found in the range between 40 and 65°C and between pH 4 and 5.5, indicating an interesting characteristic for application in the hydrolysis of lignocellulosic biomass for biofuels production. Thermostability studies of purified BGL resulted in half-lives at 37°C of 56.3 h and at 50°C of 5.4 h. These results provide support for further studies of this enzyme towards revealing its potential biotechnological applications.

β -葡萄糖苷酶(BGL)是一种对多种糖苷底物具有特异性的水解酶,是一种具有广泛生物技术应用的酶。然而,酶的性质可以根据微生物和所采用的培养程序而有所不同。因此,为了探索新型酶的潜在生物催化应用,它们的表征是必不可少的。在这项工作中,由选定的黑曲霉在固态发酵(SSF)下培养的菌株合成的BGL进行了部分纯化,并在最佳pH,温度和热稳定性方面进行了充分的表征。在色谱柱上使用manae -琼脂糖进行一步纯化,得到比活性(17.1 IU/mg蛋白质)的酶溶液,足以用于表征程序。电泳SDS-PAGE和大小排除色谱分析结果估计分子质量为60 kDa。在40至65°C和pH 4至5.5之间的范围内发现较高的酶活性,这表明了用于生产生物燃料的木质纤维素生物质水解的有趣特性。纯化BGL的热稳定性研究结果表明,在37℃和50℃的半衰期分别为56.3 h和5.4 h。这些结果为进一步研究该酶以揭示其潜在的生物技术应用提供了支持。
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引用次数: 35
Biotransformation of Indigo Pigment by Indigenously Isolated Pseudomonas sp. HAV-1 and Assessment of Its Antioxidant Property. 本土分离假单胞菌HAV-1对靛蓝色素的生物转化及其抗氧化性能评价。
Pub Date : 2014-01-01 Epub Date: 2014-11-17 DOI: 10.1155/2014/109249
Aditi Dua, Kishor Chauhan, Hilor Pathak

Chemical synthesis of indigo poses harsh environmental hazards and adverse human health effects. This necessitates an environment-friendly and producer-friendly approach for indigo production. The present study was thus significant as it reports an indigenously isolated potential indigo pigment producing culture identified as Pseudomonas sp. HAV-1 with noteworthy antioxidant property. The bioindigo pigment was characterized using various analytical techniques. The pigment production was enhanced from 412 μg mL(-1) to 700 μg mL(-1) by optimizing the growth parameters. Furthermore, the antioxidant property of indigo pigment is hitherto unexplored. This property can significantly append to its therapeutic potential. The bioindigo pigment produced by Pseudomonas sp. HAV-1 depicted 2.2 μM ascorbic acid equivalent antioxidant property. More to the point, the present work addresses a footstep towards green production of indigo.

靛蓝的化学合成对环境造成严重危害,并对人体健康造成不利影响。这就需要一种对环境友好和生产者友好的方法来生产靛蓝。因此,本研究具有重要意义,因为它报道了一种本土分离的可能产生靛蓝色素的培养物,鉴定为假单胞菌sp. HAV-1,具有显著的抗氧化性能。采用各种分析技术对生物靛蓝色素进行了表征。通过优化生长参数,使色素产量从412 μg mL(-1)提高到700 μg mL(-1)。此外,靛蓝色素的抗氧化性能迄今尚未被开发。这一特性可以显著增加其治疗潜力。假单胞菌HAV-1制备的生物靛蓝色素具有2.2 μM抗坏血酸当量的抗氧化性能。更重要的是,目前的工作是朝着绿色生产靛蓝迈出的一步。
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引用次数: 12
Algae oil: a sustainable renewable fuel of future. 藻类油:未来的可持续可再生燃料。
Pub Date : 2014-01-01 Epub Date: 2014-05-05 DOI: 10.1155/2014/272814
Monford Paul Abishek, Jay Patel, Anand Prem Rajan

A nonrenewable fuel like petroleum has been used from centuries and its usage has kept on increasing day by day. This also contributes to increased production of greenhouse gases contributing towards global issues like global warming. In order to meet environmental and economic sustainability, renewable, carbon neutral transport fuels are necessary. To meet these demands microalgae are the key source for production of biodiesel. These microalgae do produce oil from sunlight like plants but in a much more efficient manner. Biodiesel provides more environmental benefits, and being a renewable resource it has gained lot of attraction. However, the main obstacle to commercialization of biodiesel is its cost and feasibility. Biodiesel is usually used by blending with petro diesel, but it can also be used in pure form. Biodiesel is a sustainable fuel, as it is available throughout the year and can run any engine. It will satisfy the needs of the future generation to come. It will meet the demands of the future generation to come.

像石油这样的不可再生燃料已经使用了几个世纪,而且它的使用量每天都在增加。这也会增加温室气体的排放,从而导致全球变暖等全球性问题。为了满足环境和经济的可持续性,可再生的、碳中性的运输燃料是必要的。为了满足这些需求,微藻是生产生物柴油的关键来源。这些微藻确实像植物一样从阳光中产生油,但其效率要高得多。生物柴油具有更多的环境效益,作为一种可再生资源,它受到了广泛的关注。然而,生物柴油商业化的主要障碍是其成本和可行性。生物柴油通常与石油柴油混合使用,但它也可以以纯形式使用。生物柴油是一种可持续的燃料,因为它全年都可以使用,并且可以驱动任何发动机。它将满足未来一代的需要。它将满足未来一代的需求。
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引用次数: 68
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