目錄
前言
第一部分 生物質(zhì)資源的分析與檢測(cè) 1
實(shí)驗(yàn)1 植物纖維原料的全分析實(shí)驗(yàn)1
實(shí)驗(yàn)1-1 樣品的采集與制備 1
實(shí)驗(yàn)1-2 固形物含量的測(cè)定 8
實(shí)驗(yàn)1-3 灰分含量的測(cè)定 12
實(shí)驗(yàn)1-4 溶劑提取物含量的測(cè)定 17
實(shí)驗(yàn)1-5 聚糖類物質(zhì)含量的測(cè)定 23
實(shí)驗(yàn)1-6 木質(zhì)素含量的測(cè)定 30
設(shè)計(jì)型實(shí)驗(yàn) 糧食作物類原料全分析——谷物部分（大米、稻殼）和秸稈部分 35
第二部分 酶工程實(shí)驗(yàn)44
實(shí)驗(yàn)1 酶的提取與保藏 44
實(shí)驗(yàn)2 酶的物理化學(xué)性質(zhì)測(cè)定 51
實(shí)驗(yàn)2-1 酶蛋白濃度的測(cè)定 51
實(shí)驗(yàn)2-2 酶的分子質(zhì)量測(cè)定 56
實(shí)驗(yàn)2-3 濾紙酶活力的測(cè)定 65
實(shí)驗(yàn)2-4 木聚糖酶/CMC酶活力的測(cè)定 71
實(shí)驗(yàn)2-5 β-葡糖苷酶活力的測(cè)定 76
實(shí)驗(yàn)3 酶學(xué)性質(zhì)測(cè)定 84
實(shí)驗(yàn)3-1 pH對(duì)酶活性的影響 84
實(shí)驗(yàn)3-2 溫度對(duì)酶活性的影響 88
實(shí)驗(yàn)3-3 酶促反應(yīng)動(dòng)力學(xué)常數(shù)測(cè)定 92
實(shí)驗(yàn)3-4 正交法測(cè)定幾種因素對(duì)酶活性的影響 101
實(shí)驗(yàn)4 工業(yè)酶制劑的生產(chǎn)應(yīng)用 110
實(shí)驗(yàn)4-1 游離酶制劑的生產(chǎn)應(yīng)用 110
實(shí)驗(yàn)4-2 固定化酶制劑的生產(chǎn)應(yīng)用 117
實(shí)驗(yàn)4-3 高效液相色譜法測(cè)定纖維素酶水解產(chǎn)物 124
實(shí)驗(yàn)4-4 離子色譜法測(cè)定木聚糖酶水解產(chǎn)物 133
第三部分 發(fā)酵工程實(shí)驗(yàn)139
實(shí)驗(yàn)1 液態(tài)發(fā)酵 139
實(shí)驗(yàn)1-1 搖瓶液體發(fā)酵實(shí)驗(yàn)139
實(shí)驗(yàn)1-2 固定化細(xì)胞發(fā)酵實(shí)驗(yàn)145
實(shí)驗(yàn)1-3 發(fā)酵罐液體發(fā)酵實(shí)驗(yàn)151
實(shí)驗(yàn)1-4 液體發(fā)酵體系的在線檢測(cè)與控制實(shí)驗(yàn)156
實(shí)驗(yàn)1-5 發(fā)酵廢液化學(xué)需氧量的測(cè)定 161
實(shí)驗(yàn)1-6 發(fā)酵廢液生化需氧量的測(cè)定 166
實(shí)驗(yàn)1-7 米根霉發(fā)酵制備L-乳酸 173
實(shí)驗(yàn)2 固態(tài)發(fā)酵 180
實(shí)驗(yàn)2-1 固態(tài)發(fā)酵產(chǎn)纖維素酶 180
實(shí)驗(yàn)2-2 固態(tài)發(fā)酵產(chǎn)β-葡糖苷酶 187
第四部分 細(xì)胞工程實(shí)驗(yàn)192
實(shí)驗(yàn)1 植物細(xì)胞工程實(shí)驗(yàn)192
實(shí)驗(yàn)1-1 植物組織培養(yǎng)——愈傷組織培養(yǎng) 192
實(shí)驗(yàn)1-2 原生質(zhì)體的分離與融合 199
實(shí)驗(yàn)2 動(dòng)物細(xì)胞工程實(shí)驗(yàn)208
實(shí)驗(yàn)2-1 細(xì)胞的凍存和復(fù)蘇 208
實(shí)驗(yàn)2-2 動(dòng)物細(xì)胞的原代培養(yǎng)和傳代培養(yǎng) 214
第五部分 基因工程實(shí)驗(yàn)219
實(shí)驗(yàn)1 細(xì)菌基因組 DNA的提取 219
實(shí)驗(yàn)2 PCR技術(shù)擴(kuò)增目的基因 224
實(shí)驗(yàn)3 目的基因和載體的連接與轉(zhuǎn)化 229
實(shí)驗(yàn)4 重組質(zhì)粒的提取 233
實(shí)驗(yàn)5 重組質(zhì)粒的酶切與驗(yàn)證 238
第六部分 生物分離工程實(shí)驗(yàn)242
實(shí)驗(yàn)1 超聲波法破碎酵母細(xì)胞及破碎率的測(cè)定 242
實(shí)驗(yàn)2 雙水相系統(tǒng)對(duì)蛋白質(zhì)的分配測(cè)定 247
實(shí)驗(yàn)3 超濾技術(shù)分離濃縮木聚糖酶 251
實(shí)驗(yàn)4 木聚糖酶的硫酸銨分級(jí)沉淀法 257
實(shí)驗(yàn)5 蘆薈多糖的提取分離 262
實(shí)驗(yàn)6 凝膠層析法分離純化蛋白質(zhì) 267
第七部分 生物材料工程實(shí)驗(yàn)273
實(shí)驗(yàn)1 蟹殼中幾丁質(zhì)的提取與純化 273
實(shí)驗(yàn)2 幾丁質(zhì)納米分散液的制備 277
實(shí)驗(yàn)3 纖維素納米分散液的制備 280
實(shí)驗(yàn)4 幾丁質(zhì)脫乙酰化及其納米分散液的制備 283
參考文獻(xiàn)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