全书分为七个部分,每部分实验完全按照学生实验要求编写,均分列了该实验的原理、目的和所用材料，详细叙述了实验步骤，并将实验的关键点和应注意的问题列入注意事项之中，并且列出思考题，供学生复习，老师检查教学成果。全书涵盖了生物工程的五大工程，并增加信心的生物材料工程，每章节内容相对独立，但有些实验会运用到若干工程领域的实验技术，所以又相互关联。
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目录
前言
第一部分 生物质资源的分析与检测 1
实验1 植物纤维原料的全分析实验1
实验1-1 样品的采集与制备 1
实验1-2 固形物含量的测定 8
实验1-3 灰分含量的测定 12
实验1-4 溶剂提取物含量的测定 17
实验1-5 聚糖类物质含量的测定 23
实验1-6 木质素含量的测定 30
设计型实验 粮食作物类原料全分析——谷物部分（大米、稻壳）和秸秆部分 35
第二部分 酶工程实验44
实验1 酶的提取与保藏 44
实验2 酶的物理化学性质测定 51
实验2-1 酶蛋白浓度的测定 51
实验2-2 酶的分子质量测定 56
实验2-3 滤纸酶活力的测定 65
实验2-4 木聚糖酶/CMC酶活力的测定 71
实验2-5 β-葡糖苷酶活力的测定 76
实验3 酶学性质测定 84
实验3-1 pH对酶活性的影响 84
实验3-2 温度对酶活性的影响 88
实验3-3 酶促反应动力学常数测定 92
实验3-4 正交法测定几种因素对酶活性的影响 101
实验4 工业酶制剂的生产应用 110
实验4-1 游离酶制剂的生产应用 110
实验4-2 固定化酶制剂的生产应用 117
实验4-3 高效液相色谱法测定纤维素酶水解产物 124
实验4-4 离子色谱法测定木聚糖酶水解产物 133
第三部分 发酵工程实验139
实验1 液态发酵 139
实验1-1 摇瓶液体发酵实验139
实验1-2 固定化细胞发酵实验145
实验1-3 发酵罐液体发酵实验151
实验1-4 液体发酵体系的在线检测与控制实验156
实验1-5 发酵废液化学需氧量的测定 161
实验1-6 发酵废液生化需氧量的测定 166
实验1-7 米根霉发酵制备L-乳酸 173
实验2 固态发酵 180
实验2-1 固态发酵产纤维素酶 180
实验2-2 固态发酵产β-葡糖苷酶 187
第四部分 细胞工程实验192
实验1 植物细胞工程实验192
实验1-1 植物组织培养——愈伤组织培养 192
实验1-2 原生质体的分离与融合 199
实验2 动物细胞工程实验208
实验2-1 细胞的冻存和复苏 208
实验2-2 动物细胞的原代培养和传代培养 214
第五部分 基因工程实验219
实验1 细菌基因组 DNA的提取 219
实验2 PCR技术扩增目的基因 224
实验3 目的基因和载体的连接与转化 229
实验4 重组质粒的提取 233
实验5 重组质粒的酶切与验证 238
第六部分 生物分离工程实验242
实验1 超声波法破碎酵母细胞及破碎率的测定 242
实验2 双水相系统对蛋白质的分配测定 247
实验3 超滤技术分离浓缩木聚糖酶 251
实验4 木聚糖酶的硫酸铵分级沉淀法 257
实验5 芦荟多糖的提取分离 262
实验6 凝胶层析法分离纯化蛋白质 267
第七部分 生物材料工程实验273
实验1 蟹壳中几丁质的提取与纯化 273
实验2 几丁质纳米分散液的制备 277
实验3 纤维素纳米分散液的制备 280
实验4 几丁质脱乙酰化及其纳米分散液的制备 283
参考文献286
Contents
Part 1 Analysis and Test of Biomass Resources 4
Experiment 1 Full Analysis of Lignocellulose 4
Experiment 1-1 Collectionsand Preparation ofSamples for Compositional Analysis 4
Experiment 1-2 Determination of Total Solids in Biomass 10
Experiment 1-3 Determination of Ash in Biomass 14
Experiment 1-4 Quantification of the Solvent Extractives 20
Experiment 1-5 Determination of Polysaccharides 26
Experiment 1-6 Determination of Lignin Concentration 32
Designed Experiment Comprehensive Analysis of Grain Crop Materials——Grain Parts（Rice、Rice Husk）and Straw Parts 39
Part 2 Enzyme Engineering Experiments 47
Experiment 1 Extraction and Preservation of Enzyme 47
Experiment 2 Physical and Chemical Characterization of Enzyme 53
Experiment 2-1 Protein Concentration Determination of Enzyme 53
Experiment 2-2 Molecular Weight Determination of Enzyme 60
Experiment 2-3 Activity Determination of Filter Paper Enzyme 68
Experiment 2-4 Xylanase/CMC Enzyme Activity Deter-Mination 73
Experiment 2-5 Determination of β-glucosidase Activity 80
Experiment 3 Characterization of Enzyme 86
Experiment 3-1 Influence of pH on Enzyme Activity 86
Experiment 3-2 Influence of Temperature on Enzyme Activity 90
Experiment 3-3 Determination of the Michaelis Constant of Enzyme 96
Experiment 3-4 Influence of Several Factors on Enzyme Activity Determined by the Orthogonal Method 105
Experiment 4 The Production and Applications of Industrial Enzyme Agent 113
Experiment 4-1 The Production and Applications of Free Enzyme Agent 113
Experiment 4-2 The Production and Applications of Immobilized Enzyme Agent 120
Experiment 4-3 Determination of Cellulase Hydrolysis Products Using High Performance Liquid Chromatography 128
Experiment 4-4 Determination of the Products of Xylan Enzymolysis Using Anion-Exchange Chromatography 136
Part 3 Experiments for Fermentation Engineering 142
Experiment 1 Submerge Fermentation 142
Experiment 1-1 Submerged Fermentation in Shaking Flask 142
Experiment 1-2 Immobilized Cell Fermentation 148
Experiment 1-3 Submerged Fermentation in the Fermentation Tank 153
Experiment 1-4 Online Detection and Control of Submerged Fermentation System 158
Experiment 1-5 Determination of COD in Fermentation Waste 163
Experiment 1-6 Determination of Biochemical Oxygen Demand in Fermentation Waste 169
Experiment 1-7 L-lactic Acid Production by Rhizopus oryzae Fermentation 176
Experiment 2 Solid State Fermentation 183
Experiment 2-1 Cellulase Production from Solid State Fermentation 183
Experiment 2-2 β-glucosidase Production fromSolid State Fermentation 189
Part 4 Cell Engineering Experiments 195
Experiment 1 Plant Cell Engineering 195
Experiment 1-1 Plant Tissue Culture——Callus Culture 195
Experiment 1-2 Protoplast Isolation and Fusion 203
Experiment 2 Animal Cell Engineering 211
Experiment 2-1 Cell Cryopreservation and Recovery 211
Experiment 2-2 Primary Culture and Subculture of Animal Cells 216
Part 5 Genetic Engineering Experiments 221
Experiment 1 The Extraction of the Bacteria Genome DNA 221
Experiment 2 Target Gene Amplification with PCR Technique 226
Experiment 3 The Ligation and Transformation of Target Genes and Vectors 231
Experiment 4 Isolation of Recombinant Plasmids 235
Experiment 5 Enzyme Digestion and Verification of Recombinant Plasmid 240
Part 6 Bioseparation Engineering Experiments 244
Experiment 1 Ultrasonic Disruption of Yeast Cells and Determination of Disruption Rate 244
Experiment 2 Distribution Determination of Proteins in Aqueous Two-phase System 249
Experiment 3 Separate and Concentrate Xylanase Using Ultrafiltration 254
Experiment 4 The Purification of Xylanase by Gradient Concentrations Precipitation of Ammonium Sulfate 259
Experiment 5 The Extraction and Purification of the Aloe Polysaccharides 264
Experiment 6 Purification of Proteins by Gel Filtration Chromatography Method 270
Part 7 Biomaterial Engineering Experiments 275
Experiment 1 Extraction and Purification of Chitin from Crab Shells 275
Experiment 2 Preparation of Aqueous Chitin Nanofiber Dispersion 278
Experiment 3 Preparation of Aqueous Cellulose Nanofiber Dispersion 281
Experiment 4 Deacetylation of Chitin and Preparation of Nanofiber Dispersion 284
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