基于生产工艺管理的生物工程综合性实验设计

生物工程综合性实验课程以企业人才需求为导向,解决实际生产过程中的复杂工程问题为教学目标,利用两步酶转化法制备L-天冬氨酸和L-丙氨酸的工艺路线,结合生物工程专业生产工艺管理的特点,借鉴生产企业现场管理经验,实施四班三运转的实验运行方式。成绩考核加入交接班总结评价与团队协作评价,该课程设置了包含多门专业核心课程原理、方法与实验技术和企业生产管理模式的新型生物工程综合性实验教学内容,并通过教学实践,持续改进,形成完整的实验教学过程与考核机制。生物工程综合性实验课程取得了良好的教学效果,促进了生物工程专业实验教学的发展。     

基于正交试验法的景观水体叶绿素a 最佳提取条件研究

根据水体初级生产力测定常用方法中叶绿素a 测定法, 采用3 因素3 水平的正交法设计试验((L9(34)), 综合探究某喷泉水池景观水体测定过程中滤膜处理条件、浸提试剂和浸提温度等多因素多水平的共同作用下叶绿素a 提取效果的差异, 改进的正交试验设计规避了单因素控制变量实验的片面性。试验结果表明该喷泉池水的叶绿素a 含量较高,参考湖库景观水体富营养化分级标准已达到富营养化水平。提取过程中不同的正交试验组结果存在差异, 其中冷冻处理滤膜、丙酮乙醇混合溶剂、4 ℃条件下提取值最高, 为62.3 μg⋅L− 1。上述景观水体叶绿素a 提取条件组合(A2B3C1) 为正交试验最优水平组合, 采用正交试验的方法探究叶绿素a 提取最佳条件有利于对传统实验方法改进。     

大苞萱草ISSR-PCR反应体系的建立与优化

分别采用正交设计和单因子试验两种方法对影响大苞萱草ISSR-PCR反应体系的4个因素(Taq酶,Mg2+,dNTP,引物)在3个水平上进行优化试验。正交设计选用L9(34)方案,采用直观分析法获得影响因素最佳反应水平。单因子试验分别研究各因素对ISSR-PCR反应的影响情况,找出最佳反应水平。两种方法所得影响因素最佳水平存在差异,通过综合比较与分析,最终建立了大苞萱草ISSR-PCR的最佳反应体系。在20μL的反应体系中:Taq酶1 U,Mg2+1.5 mmol/L,dNTP0.20 mmol/L,引物0.4μmol/L,1×PCR buffer,30 ng模板DNA。在此基础上筛选出10个扩增稳定、多态性丰富的ISSR引物,并通过梯度PCR试验,确定引物最佳退火温度。     

“新工科”背景下“生物化工大实验”课程教学设计与实践

“生物化工大实验”是湖南科技学院生物工程专业集中实践教学环节的必修课程,是在“新工科”背景下为提高本专业学生综合素质而开设的实验课程。课程实验项目为综合性实验及设计性实验,其内容主要结合了永州市地方特色资源、研究平台和工科人才特征合理编制而成。授课过程中,综合运用符合课程特点的启发式教学、科研案例导入式教学和互动式教学等方法,提高学生学习的兴趣,激发学生的创新能力和实践应用能力。通过课程考察和课后调查发现,生物工程专业学生的知识迁移和应用能力明显提升,学科竞赛和项目立项成绩优异。因此,基于对本课程授课内容和方法的实践及不断完善,课程教学质量得到了全面提高,为培养生物工程专业高水平创新型、应用型工科人才夯实了基础。     

组正交超饱和设计在发酵罐手工清洗方法开发中的应用

目的 开发一种适用于发酵罐手工清洗方法.方法 采用组正交超饱和设计方法(15个因素2个水平),试验因素包括9个实际影响发酵罐清洗因素和6个假因素;评价指标为发酵罐清洗后淋洗水电导率与总有机碳(total organic carbon,TOC)含量;通过标准最小二乘法拟合试验结果获得较优清洗条件,从而建立发酵罐手工清洗方...     

利用正交设计优化异色瓢虫SRAP-PCR反应体系

利用正交设计L16(45)对异色瓢虫Harmonia axyridis(Pallas)SRAP-PCR反应体系的5个因素(Taq酶、Mg2+、模板DNA、dNTPs、引物)在4个水平上进行优化试验,试验结果用DPS和MINITAB软件进行分析,建立了异色瓢虫SRAP-PCR反应的最佳体系,即在20μL体系中模板50~100ng、引物0.25μmol/L、dNTPs0.1mmol/L、Taq DNA聚合酶1.5U、Mg2+0.375~0.625mmol/L。并对反应体系进行梯度退火试验,得到最佳退火温度为50.3℃。这一优化系统的建立,为今后利用SRAP技术进行瓢虫遗传图谱的构建、多态性分析和基因定位奠定了技术基础。     

黑曲霉纤维素酶高产固态培养基的快速优选研究

研究采用正交设计,对影响黑曲霉 Ｆ_２７产纤维素酶的 １０个营养因素在较少水平下进行初选,从中选出对产酶影响显著的４个因素．在此基础上采用均匀设计,在较多水平下进行较少次数的试验,将试验结果用微机逐步回归处理,得到最优回归方程,由回归方程计算出预测极值点和预测极值、优化出高产纤维素酶固态培养基配方．经试验验证,试验实测值和预测值基本相符, ＣＭＣａｓｅ活力平均达 ３７０５Ｕ／ｇ, ＦＰＡ活力平均达 ７．４Ｕ／ｇ．表明对于多因子试验,应用正交设计结合均匀设计的试验方法是快速、简便、可行的．     

黑木耳ISSR-PCR反应体系的正交优化

采用正交试验设计的方法,对黑木耳ISSR-PCR(简单重复序列区间-多聚酶链反应)反应体系中的5种主要因素(Taq聚合酶,Mg2+,模板DNA,dNTP及引物)4个水平进行优化筛选,确立了适合黑木耳ISSR分析的优化反应体系(20μL),通过梯度PCR试验筛选得到相应引物的最佳退火温度。     

应用降落PCR和正交设计优化AtSUC9 PCR反应体系

目的:建立最适的AtSUC9 PCR反应体系。方法:应用降落PCR和正交设计,从Mg2 、dNTPs、引物、甘油4种因素3个水平,对拟南芥AtSUC99 PCR反应体系进行优化。结果:确立了适合拟南芥AtSUC99 PCR反应体系:在20μl反应体系中,含模板1μg,引物(20μmol/L)0.5μl,dNTPs(10mmol/L)0.3μl,10×PCRbuffer2μl,Mg2 (25mmol/L)1.2μl,TaqDNAPolymerase1U,甘油10μl。结论:降落PCR和正交设计是一种有效的植物基因PCR体系优化方法。     

生物工程类课程“半自助式”教学模式的探索与实践

介绍“半自助式”教学模式提出的背景、涵义和研究理念,明晰该模式的实施方案和考核评分体系.以《生物工艺学》课程的教学组织为典型案例,分四个阶段介绍如何通过实践题目的布置引出课程内容体系,如何安排基础知识学习,如何将基础知识应用于具体生物工程产品生产以及如何做生产工艺的工厂设计,分析成效,以期为生物工程类课程教学提供一种新型教学模式,提高学生工程实践能力.     

离子束生物工程装置剂量测量系统开发

介绍了离子束生物工程装置(ion beam bio-engineering device,IBBe-Device)中生物样品离子注入剂量测量原理,并根据回推热测法设计出新的剂量测量方案。整个系统由测量靶和温度传感器、数据采集、计算机数据处理软件以及控制系统组成,实现计算机测量系统的计算机控制,并达到比较理想的测量结果。最后讨论了进一步改进IBBe-Device剂量测量系统的方法。     

Experimental design methods for bioengineering applications

Experimental design is a form of process analysis in which certain factors are selected to obtain the desired responses of interest. It may also be used for the determination of the effects of various independent factors on a dependent factor. The bioengineering discipline includes many different areas of scientific interest, and each study area is affected and governed by many different factors. Briefly analyzing the important factors and selecting an experimental design for optimization are very effective tools for the design of any bioprocess under question. This review summarizes experimental design methods that can be used to investigate various factors relating to bioengineering processes. The experimental methods generally used in bioengineering are as follows: full factorial design, fractional factorial design, Plackett–Burman design, Taguchi design, Box–Behnken design and central composite design. These design methods are briefly introduced, and then the application of these design methods to study different bioengineering processes is analyzed.     

dsDNA芯片的DNA结合蛋白质计算机模拟系统

dsDNA芯片(double-stranded DNA microarray)是用于DNA结合蛋白质表达研究的新技术,应用一种新的dsDNA芯片制作方法,研究了dsDNA探针设计,开发了相应的计算机模拟系统—DBP软件。可实现芯片上dsDNA探针的选取以及DNA结合蛋白质检测的实验过程模拟,功能包括模拟酶切、模拟电泳、模拟杂交等实验环节。DBP系统包括了可以不断完善的DNA结合蛋白质数据库。对研究dsDNA芯片,检测和分析在生物 体中DNA结合蛋白质的表达谱,揭示细胞的分化和凋亡、信号转导、DNA转录调控的分子机制有重要意义。     

数理统计方法优化单细胞蛋白发酵培养基研究

采用正交试验和中心组合设计相结合的数理统计方法,在２Ｌ发酵罐中对紫云英汁液培养单细胞蛋白发酵培养基进行筛选、优化,试验结果表明较优培养基为：初糖２．３％,酵母粉０．１６％,ＫＨ_２ＰＯ_４０．２％,ＭｇＳＯ_４０．０５％。酵母浓度最高达１０．４６ｇ／Ｌ,基质生长得率为０．５５ｇ菌体／ｇ基质,基质转化率为８３％。研究结果同时说明采用数理统计方法设计实验,工作量小,效率高,结果准确。     

Taguchi approach significantly increases bioremediation process efficiency: a case study with Hg (II) removal by Pseudomonas aeruginosa

AIM: Optimization of process parameters for mercury removal by an Hg (II)-reducing Pseudomonas aeruginosa strain. METHODS AND RESULTS: A strain of Ps. aeruginosa was found to reduce 10 mg l(-1) Hg (II) to Hg0 with 70% efficiency in 24 h. To optimize process performance, a statistical tool--Taguchi design of experiments (DOE)--was used to carry out 18 well-defined experiments (L18 Orthogonal array) with eight variable parameters (viz. agitation, temperature, pH, carbon source, medium volume: flask volume ratio and concentrations of Hg (II), ammonium sulfate and yeast extract). When data obtained were analyzed using specialized software for Taguchi design, Qualitek-4 (Nutek Inc., MI, USA), Hg (II) reduction efficiency was predicted to be 95% in 24 h under the optimized process parameters (also suggested by the software). In the validation experiment, Hg (II) removal of 99.29% in 24 h was indeed obtained. CONCLUSIONS: Using Taguchi DOE, Hg (II) reduction (and hence its removal) using Ps. aeruginosa could be improved by 29.3%. SIGNIFICANCE AND IMPACT OF THE STUDY: Taguchi approach could be employed as an efficient and time-saving strategy for parameter optimization in bioremediation processes.     

Development of a target component extraction method from GC-MS data with an in-house program for metabolite profiling

After gas chromatography-mass spectrometry (GC-MS) analysis, data processing, including retention time correction, spectral deconvolution, peak alignment, and normalization prior to statistical analysis, is an important step in metabolomics. Several commercial or free software packages have been introduced for data processing, but most of them are vendor dependent. To design a simple method for Agilent GC/MS data processing, we developed an in-house program, "CompExtractor", using Microsoft Visual Basic. We tailored the macro modules of an Agilent Chemstation and implanted them in the program. To verify the performance of CompExtractor processing, 30 samples from the three species of the genus Papaver were analyzed with Agilent 5973 MSD GC-MS. The results of CompExtractor processing were compared with those of AMDIS-SpectConnect processing by hierarchical cluster analysis (HCA) and principal component analysis (PCA). The two methods showed good classification according to their species in HCA. The PC1+PC2 scores were 54.32-63.62% for AMDIS-SpectConnect and 56.65-85.92% for CompExtractor in PCA. Although the CompExtractor processing method is an Agilent GC-MS-specific application and the target compounds must be selected first, it can extract the target compounds more precisely in the raw data file with batch mode and simultaneously assemble the matrix text file.     

生物信息学在基因芯片中的应用

生物信息学和基因芯片是生命科学研究领域中的两种新方法和新技术,生物信息学与基因芯片密切相关,生物信息学促进了基因芯片的研究与应用,而基因芯片则丰富了生物信息学的研究内容。本论文探讨生物信息学在基因芯片中的应用,将生物信息学方法运用到高密度基因芯片设计和芯片实验数据管理及分析。从信息学的角度提出基因芯片设计准则,提出寡核苷酸探针的优化设计方法,将该方法运用于再测序型芯片和基因表达型芯片的设计,在此基础上研制出高密度基因芯片设计软件系统和实验结果分析系统。     

A survey of computational and physical methods applied to solid-state fermentation

During the last decade, significant effort has been made to apply computational and physical methods to solid-state fermentation (SSF). This had positive impact both on our understanding of the basic principles underlying this old technology, and on the latest progress made in industrial bioengineering. Guidelines on bioreactor design and operation including scale-up, new methods for biomonitoring and advanced control strategies are among the most important outcomes of practical use. Nevertheless, there still is a lack of experimental data, which hampers parameter identification and thus broader use of mathematical modeling. More attention should therefore be paid to combining and concentrating modern physical techniques and computational approaches in order to allow better model validation and thus further progress in rational bioengineering of SSF.     

Orthogonal array design in optimizing ERIC-PCR system for fingerprinting rat's intestinal microflora

AIMS: The aim of the present study was to rapidly optimize enterobacterial repetitive intergenic consensus (ERIC)-PCR amplification systems for fingerprinting rat's intestinal microflora. METHODS AND RESULTS: Orthogonal array design and statistic analysis methods were attempted to rapidly optimize ERIC-PCR reaction system for fingerprinting intestinal microflora. The results showed that variations of the four factors (Mg(2+), dNTP, primer and HotstarTaq polymerase concentrations) changed the fingerprinting patterns significantly. The order of effects of those factors on fingerprinting patterns was primers (F = 274.000, P = 0.000), Hotstar Taq polymerase (F = 197.000, P = 0.001), Mg(2+) (F = 181.000, P = 0.001) and dNTP (F = 27.000, P = 0.011). The optimal ERIC-PCR condition was containing 200 micromol l(-1) dNTP, 2.5 mmol l(-1) Mg(2+), 0.4 micromol l(-1) primer, 1 U HotstarTaq DNA polymerase namely 25 microl reaction system, which is proved to be a simple, fast and reliable method suitable for fingerprinting rat's intestinal microflora. CONCLUSIONS: The results suggest that Mg(2+), dNTP, primer and HotstarTaq polymerase concentrations play important roles on ERIC-PCR fingerprinting patterns. Orthogonal array design is a considerable method to optimize ERIC-PCR reaction system for its rapidness, simplicity, potential to investigate mutual effects of parameters. SIGNIFICANCE AND IMPACT OF THE STUDY: It is the first report on optimization of ERIC-PCR amplification systems for fingerprinting intestinal microflora using orthogonal array design or statistic analysis methods and systematically observing the effects of variables of reaction conditions.     

氨基酸的冷电弧-光催化-吸附脱色工艺优化

将TiO2光催化剂负载于活性炭,然后将活性炭装填于冷电弧-光催化-吸附集成反应器中,用该反应器对豆粕氨基酸进行脱色。以豆粕氨基酸的脱色率和氨基酸保留率为指标,探讨处理电压、处理时间、溶液pH和活性炭填充量4个因素对脱色效果的影响。在单因素试验的基础上,利用二次回归正交旋转组合设计建立回归模型方程确定最佳工艺条件,并对方程进行回归分析和系数检验。结果表明:回归方程在#=0.01水平上高度显著。各因素对脱色效果的影响程度大小依次为:溶液pH>活性炭填充量>处理时间>处理电压。最佳脱色条件为处理电压20kV、处理时间11min、溶液pH 3.6和活性炭填充量63g,此条件下氨基酸溶液脱色率为94.8%,保留率为83.1%。