Abstract:

Abstract:This article summarized the reviews and research articles published in Chinese Journal of Biotechnology in the field of biomanufacturing in 2021.The article covered major chassis cells such as Escherichia coli,Bacillus subtilis,Corynebacterium glutamicum,Saccharomyces cerevisiae,filamentous fungi,non-model bacteria and non-conventional yeasts.Moreover,this article summarized the advances in the production of amino acids,organic acids,vitamins,higher alcohols,natural compounds (terpenoids,flavonoids,alkaloids),antibiotics,enzymes and enzyme-catalyzed products,biopolymers,as well as the utilization of biomass and one-carbon materials.The key technologies used in the construction of cell factories,such as regulation,evolution,and high-throughput screening,were also included.This article may help the readers better understand the R & D trend in biomanufacturing driven by engineered microbes.

Abstract:Unnatural amino acids are widely used in medicine,pesticide,material,and other industries and the green and efficient synthesis has attracted a lot of attention.In recent years,with the rapid development of synthetic biology,microbial cell factories have become a promising means for biosynthesis of unnatural amino acids.This study reviewed the construction and application of microbial cell factories for unnatural amino acid,including the synthetic pathway reconstruction,design/modification of key enzymes and their coordinated regulation with precursors,blocking of competitive alternative pathways,and construction of cofactor circulation systems.Meanwhile,on the basis of the new principles for designing the microbial cell factories,new biosynthetic pathways adapted to cells and the production environment,as well as new biomanufacturing system established based on cell adaptive evolution and intelligent fermentation regulation,we looked forward to the further construction and application of microbial cell factories for industrial bio-production.

Abstract:Tetrapyrrole compounds are a class of compounds with important functions.They exist in living organisms and have been widely used in agriculture,food,medicine,and other fields.The cumbersome process and high cost of chemical synthesis,as well as the shortcomings of unstable quality of animal and plant extraction methods,greatly hampered the industrial production and applications of tetrapyrrole compounds.In recent years,the rapid development of synthetic biology has provided new tools for microorganisms to efficiently synthesize tetrapyrrole compounds from renewable biomass resources.This article summarizes various strategies for the biosynthesis of tetrapyrrole compounds,discusses methods to improve its biosynthesis efficiency and future prospects,with the aim to facilitate the research on biosynthesis of tetrapyrrole compounds.

Abstract:Aerobic methane oxidizing bacteria (methanotrophs) can use methane as carbon source and energy source,eliminating 10%-20% of global methane.Methanotrophs can also effectively synthesize valuable methane-derived products.This article introduced the methane oxidizing mechanism of methanotrophs,and summarized the practical application and research hotspots of methanotrophs in the field of methane emission reduction in the landfill,ventilation air methane mitigation in coal mines,valuable chemicals biosynthesis,as well as oil and gas reservoir exploration.Main factors influencing the pollutant removal and the biosynthesis efficiency in various applications were also discussed.Based on the study of large-scale cultivation of methanotrophs,some measures to benefit the application and promotion of aerobic methane oxidizing biotechnology were proposed.This includes investigating the effect of intermediate metabolites on methanotrophs activity and population structure,and exploiting economical and efficient alternative culture media and culture techniques.

Abstract:Human activities increase the concentration of atmospheric carbon dioxide (CO₂),which leads to global climate warming.Microbial CO₂ fixation is a promising green approach for carbon neutral.In contrast to autotrophic microorganisms,heterotrophic microorganisms are characterized by fast growth and ease of genetic modification,but the efficiency of CO₂ fixation is still limited.In the past decade,synthetic biology-based enhancement of heterotrophic CO₂ fixation has drawn wide attention,including the optimization of energy supply,modification of carboxylation pathway,and heterotrophic microorganisms-based indirect CO₂ fixation.This review focuses on the research progress in CO₂ fixation by heterotrophic microorganisms,which is expected to serve as a reference for peaking CO₂ emission and achieving carbon neutral by microbial CO₂ fixation.

Abstract:Anaerobic ammonia oxidation (ANAMMOX) process is an efficient and low-cost biological nitrogen removal process.However,it still faces some challenges in mainstream applications due to the limitation of substrate types and nitrate accumulation.In recent years,the combined process of anammox has been widely studied to solve the above problems.In this paper,the combined processes of anammox developed in recent years are reviewed,and discussed from the process principle,advantages and disadvantages,influencing factors,process extensibility and the key bottlenecks existing in the promotion and application,as well as the relevant work of the subject group.Finally,we take an outlook on the development of the combined anaerobic ammonia oxidation process in municipal domestic wastewater treatment.

Abstract:Yarrowia lipolytica is a non-conventional yeast with unique physiological and metabolic characteristics.It is suitable for production of various products due to its natural ability to utilize a variety of inexpensive carbon sources,excellent tolerance to low pH,and strong ability to secrete metabolites.Currently,Y.lipolytica has been demonstrated to produce a wide range of carboxylic acids with high efficiency.This article summarized the progress in engineering Y.lipolytica to produce various carboxylic acids by using metabolic engineering and synthetic biology approaches.The current bottlenecks and solutions for high-level production of carboxylic acids by engineered Y.lipolytica were also discussed,with the aim to provide useful information for relevant studies in this field.

Abstract:The production efficiency of microbial cell factory is determined by the growth performance,product synthetic capacity,and stress resistance of the strain.Strengthening the stress resistance is the key point to improve the production efficiency of microbial cell factory.Tolerance engineering is based on the response mechanism of microbial cell factory to resist stress.Specifically,it consolidates the cell wall-cell membrane barrier to enhance the defense against stress,accelerates the stress response to improve the damage repair,and creates tolerance evolutionary tools to screen industrial microorganisms with enhanced robustness.We summarize the regulation strategies and forecast the prospects of tolerance engineering,which plays an important role in the microbial cell factories for sustainable production of natural products and bulk chemicals.

Abstract:It is among the goals in metabolic engineering to construct microbial cell factories producing high-yield and high value-added target products,and an important solution is to design efficient synthetic pathway for the target products.However,due to the difference in metabolic capacity among microbial chassises,the available substrate and the yielded products are limited.Therefore,it is urgent to design related metabolic pathways to improve the production capacity.Existing metabolic engineering approaches to designing heterologous pathways are mainly based on biological experience,which are inefficient.Moreover,the yielded results are in no way comprehensive.However,systems biology provides new methods for heterologous pathway design,particularly the graph-based and constraint-based methods.Based on the databases containing rich metabolism information,they search for and uncover possible metabolic pathways with designated strategy (graph-based method) or algorithm (constraint-based method) and then screen out the optimal pathway to guide the modification of strains.In this paper,we reviewed the databases and algorithms for pathway design,and the applications in metabolic engineering and discussed the strengths and weaknesses of existing algorithms in practical application,hoping to provide a reference for the selection of optimal methods for the design of product synthesis pathway.

Abstract:Ergothioneine is a multifunctional physiological cytoprotector,with broad application in foods,beverage,medicine,cosmetics and so on.Biosynthesis is an increasingly favored method in the production of ergothioneine.This paper summarizes the new progress in the identification of key pathways,the mining of key enzymes,and the development of natural edible mushroom species and high-yield engineering strains for ergothioneine biosynthesis in recent years.Through this review,we aim to reveal the molecular mechanism of ergothioneine biosynthesis and then employ the methods of fermentation engineering,metabolic engineering,and synthetic biology to greatly increase the yield of ergothioneine.

Abstract:Traditional methods of microbial synthesis usually rely on a single engineered strain to synthesize the target product through metabolic engineering.The key cofactors,precursors and energy are produced by the introduced complex synthetic pathways.This would increase the physiological burden of engineering strains,resulting in a decrease in the yield of target products.The modular co-culture engineering has become an attractive solution for effective heterologous biosynthesis,where product yield can be greatly improved.In the modular co-culture engineering,the coordination between the population of different modules is essential for increasing the production efficiency.This article summarized recent advances in the application of modular co-culture engineering and population control strategies.

Abstract:Bacterial multi-drug resistance (MDR) is a global challenge in the fields of medicine and health,agriculture and fishery,ecology and environment.The cross-region spread of antibiotic resistance genes (ARGs) among different species is one of the main cause of bacterial MDR.However,there is no effective strategies for addressing the intensifying bacterial MDR.The CRISPR-Cas system,consisting of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR associated proteins,can targetedly degrade exogenous nucleic acids,thus exhibiting high application potential in preventing and controlling bacterial MDR caused by ARGs.This review briefly introduced the working mechanism of CRISPR-Cas systems,followed by discussing recent advances in reducing ARGs by CRISPR-Cas systems delivered through mediators (e.g.plasmids,bacteriophages and nanoparticle).Moreover,the trends of this research field were envisioned,providing a new perspective on preventing and controlling MDR.

Abstract:Gene editing technology can be used to modify the genome of Escherichia coli for the investigation of gene functions,or to change the metabolic pathways for the efficient production of high-value products in engineered strains with genetic stability.A variety of gene editing technologies have been applied in prokaryotes,such as λ-Red homologous recombination and CRISPR/Cas9.As a traditional gene editing technique,λ-Red recombination is widely used.However,it has a few shortcomings,such as the limited integration efficiency by the integrated fragment size,the cumbersome gene editing process,and the FRT scar in the genome after recombination.CRISPR/Cas9 is widely used for genome editing at specific sites,which requires specific DNA segments according to the editing site.As the understanding of the two technologies deepens,a variety of composite gene editing techniques have been developed,such as the application of λ-Red homologous recombination in combination with homing endonucleaseⅠ-SceⅠ or CRISPR/Cas9.In this review,we summarized the basic principles of common gene editing techniques and composite gene editing techniques,as well as their applications in Escherichia coli,which can provide a basis for the selection of gene editing methods in prokaryotes.

Abstract:Extracellular vesicles (EVs),also known as membrane vesicles,are vesicular bodies secreted by eukaryotic cells and bacteria.EVs can carry proteins,DNA,RNA,and various metabolites for the exchange and transmission of substances between cells.They play contents-dependent physiological functions,such as delivering nutrients,participating in immune response,and treating cancers.Currently,most studies focus on the exploration of vesicles secreted by eukaryotic cells and gram-negative bacteria,while few studies focus on gram-positive bacteria.This review summarized the production,content composition,physiological function,and engineering of EVs secreted by gram-positive bacteria,and prospected future perspectives in this area.

Abstract:The diverse thermophilic strains of Thermoanaerobacter,serving as unique platforms with a broad range of application in biofuels and chemicals,have received wide attention from scholars and practitioners.Although biochemical experiments and genome sequences have been reported for a variety of Thermoanaerobacter strains,an efficient genetic manipulation system remains to be established for revealing the biosynthetic pathways of Thermoanaerobacter.In line with this demand,the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) systems for editing,regulating and targeting genomes have been well developed in thermophiles.Here,we reviewed and discussed the current status,associated challenges,and future perspectives of the construction of thermostable CRISPR/Cas9 genome editing systems for some representative Thermoanaerobacter species.The establishment,optimization,and application of thermostable CRISPR/Cas genome editing systems would potentially provide a foundation for further genetic modification of thermophilic bacteria.

Abstract:Intrinsically disordered proteins (IDPs) are proteins or protein regions that fail to get folded into definite three-dimensional structures but participate in various biological processes and perform specific functions.Defying the traditional protein "sequence-structure-function" paradigm,they enrich the protein "structure-function" diversity.Ubiquitous in organisms,they show extreme hydrophilicity,charged amino acids,and highly repetitive amino acid sequences,with simple arrangement.As a result,they feature highly variable binding affinities and high coordination,which facilitate their functions.IDPs play an important role in cell stress response,which can improve the tolerance to a variety of stresses,such as freezing,high salt,heat shock,and desiccation.In this study,we briefed the characteristics,classifications,and identification of IDPs,summarized the molecular mechanism in improving cell stress resistance,and described the potential applications.

Abstract:In order to explore the effect of peptidoglycan hydrolase on the viable cell counts of Bacillus amyloliquefaciens and the yield of alkaline protease,five peptidoglycan hydrolase genes (lytC,lytD,lytE,lytF and lytG) of B.amyloliquefaciens TCCC111018 were knocked out individually.The viable cell counts of the bacteria and their alkaline protease activities before and after gene deletion were determined.The viable cell counts of the knockout mutants BA ΔlytC and BA ΔlytE achieved 1.67×10⁶ CFU/mL and 1.44×10⁶ CFU/mL respectively after cultivation for 60 h,which were 32.5% and 14.3% higher than that of the control strain BA Δupp.Their alkaline protease activities reached 20 264 U/mL and 17 265 U/mL,respectively,which were 43.1% and 27.3% higher than that of the control strain.The results showed that deleting some of the peptidoglycan hydrolase genes effectively maintained the viable cell counts of bacteria and increased the activity of extracellular enzymes,which may provide a new idea for optimization of the microbial host for production of industrial enzymes.

Abstract:Covalently anchoring of a ligand/metal via polar amino acid side chain (s) is often observed in metalloenzyme,while the substitutability of metal-binding sites remains elusive.In this study,we utilized a zinc-dependent alcohol dehydrogenase from Thermoanaerobacter brockii (TbSADH) as a model enzyme,analyzed the sequence conservation of the three residues Cys37,His59,and Asp150 that bind the zinc ion,and constructed the mutant library.After experimental validation,three out of 224 clones,which showed comparative conversion and ee values as the wild-type enzyme in the asymmetric reduction of the model substrate tetrahydrofuran-3-one,were screened out.The results reveal that the metal-binding sites in TbSADH are substitutable without tradeoff in activity and stereoselectivity,which lay a foundation for designing ADH-catalyzed new reactions via metal ion replacement.

Abstract:Halomonas can grow on diverse carbon sources.As it can be used for unsterile fermentation under high-salt conditions,it has been applied as a chassis for next-generation industrial biotechnology.Short-chain volatile fatty acids,including acetate,propionate,and butyrate,can be prepared from biomass and are expected to be novel carbon sources for microbial fermentation.Halomonas sp.TD01 and TD08 were subjected to shaking culture with 10-50 g/L butyrate,and they were found to effectively synthesize poly-3-hydroxybutyrate with butyrate as the carbon source.The highest yield of poly-3-hydroxybutyrate was achieved at butyrate concentration of 20 g/L (9.12 g/L and 7.37 g/L,respectively).Butyrate at the concentration>20 g/L inhibited cell growth,and the yield of poly-3-hydroxybutyrate decreased to<4 g/L when butyrate concentration was 50 g/L.Moreover,Halomonas sp.TD08 can accumulate the copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate by using propionate and butyrate as carbon sources.However,propionate was toxic to cells.To be specific,when 2 g/L propionate and 20 g/L butyrate were simultaneously provided,cell dry weight and polymer titer were 0.83 g/L and 0.15 g/L,respectively.The addition of glycerol significantly improved cell growth and boosted the copolymer titer to 3.95 g/L,with 3-hydroxyvalerate monomer content of 8.76 mol%.Short-chain volatile fatty acids would be promising carbon sources for the production of polyhydroxyalkanoates by Halomonas.

Abstract:Proteus mirabilis lipase (PML) features tolerance to organic solvents and great potential for biodiesel synthesis.However,the thermal stability of the enzyme needs to be improved before it can be used industrially.Various computational design strategies are emerging methods for the modification of enzyme thermal stability.In this paper,the complementary algorithm-based ABACUS,PROSS,and FoldX were employed for positive selection of PML mutations,and their pairwise intersections were further subjected to negative selection by PSSM and G_REMLIN to narrow the mutation library.Thereby,18 potential single-point mutants were screened out.According to experimental verification,7 mutants had melting temperature (T_m) improved,and the ΔT_m of K208G and G206D was the highest,which was 3.75℃ and 3.21℃,respectively.Five mutants with activity higher than the wild type (WT) were selected for combination by greedy accumulation.Finally,the T_m of the five-point combination mutant M10 increased by 10.63℃,and the relative activity was 140% that of the WT.K208G and G206D exhibited certain epistasis during the combination,which made a major contribution to the improvement of the thermal stability of M10.Molecular dynamics simulation indicated that new forces were generated at and around the mutation sites,and the rearrangement of forces near G206D/K208G might stabilize the Ca²⁺ binding site which played a key role in the stabilization of PML.This study provides an efficient and user-friendly computational design scheme for the thermal stability modification of natural enzymes and lays a foundation for the modification of PML and the expansion of its industrial applications.

Abstract:Graph-theory-based pathway analysis is a commonly used method for pathway searching in genome-scale metabolic networks.However,such searching often results in many pathways biologically infeasible due to the presence of currency metabolites (e.g.H⁺,H₂O,CO₂,ATP etc.).Several methods have been proposed to address the problem but up to now there is no well-recognized methods for processing the currency metabolites.In this study,we proposed a new method based on the function of currency metabolites for transferring of functional groups such as phosphate.We processed most currency metabolites as pairs rather than individual metabolites,and ranked the pairs based on their importance in transferring functional groups,in order to make sure at least one main metabolite link exists for any reaction.The whole process can be done automatically by programming.Comparison with existing approaches indicates that more biologically infeasible pathways were removed by our method and the calculated pathways were more reliable,which may facilitate the graph-theory-based pathway design and visualization.

Abstract:8-prenylnaringenin (8-PN) is a potent estrogen with high medicinal values.It also serves as an important precursor for many prenylated flavonoids.Microbial synthesis of 8-PN is mainly hindered by the low catalytic activity of prenyltransferases (PTS) and insufficient supply of precursors.In this work,a SfN8DT-1 from Sophora flavescens was used to improve the efficiency of (2S)-naringenin prenylation.The predicted structure of SfN8DT-1 showed that its main body is comprised of 9 α-helices and 8 loops,along with a long side chain formed by nearly 120 amino acids.SfN8DT-1 mutants with different side-chain truncated were tested in Saccharomyces cerevisiae.A mutant expressing the truncated enzyme at K62 site,designated as SfND8T-1-t62,produced the highest 8-PN titer.Molecular docking of SfN8DT-1-t62 with (2S)-naringenin and dimethylallyl diphosphate (DMAPP) showed that K185 was a potentially crucial residue.Alanine scanning within a range of 0.5 nm around these two substrates showed that the mutant K185A may decrease its affinity to substrates,which also indicated K185 was a potentially critical residue.Besides,the mutant K185W enhanced the affinity to ligands implied by the simulated saturation mutation,while the saturated mutation of K185 showed a great decrease in 8-PN production,indicating K185 is vital for the activity of SfN8DT-1.Subsequently,overexpressing the key genes of Mevalonate (MVA) pathway further improved the titer of 8-PN to 31.31 mg/L,which indicated that DMAPP supply is also a limiting factor for 8-PN synthesis.Finally,44.92 mg/L of 8-PN was produced in a 5 L bioreactor after 120 h,which is the highest 8-PN titer reported to date.

Abstract:In order to overcome the challenges of insufficient restriction enzyme sites,and construct a fusion-expression vector with flexible fusion direction,we designed an LB cloning system based on the type IIS and type IIT restriction enzymes LguⅠ and BbvCⅠ.The LB cloning system is constructed by inserting the LB fragment (GCTCTTCCTCAGC) into the multiple cloning site region of the broad-host plasmid pBBR1MCS-3 using PCR.The LB fragment contains partially overlapped recognition sites of LguⅠ and BbvCⅠ.Therefore,the same non-palindromic sequence will be generated by these two restriction endonucleases digestion.This feature can be used to quickly and flexibly insert multiple genes into the expression vector in a stepwise and directed way.In order to verify the efficacy of the cloning system,two glycosyltransferase genes welB and welK of Sphingomonas sp.WG were consecutively fused to the LB cloning vector,and the recombinant plasmid was transferred into Sphingomonas sp.WG by triparental mating.The results showed that gene fusion expression has little effect on sphingan titer,but enhanced the viscosity of sphingan.The viscosity of the sphingan produced by recombinant strain Sphingomonas sp.WG/pBBR1MCS-3-LB-welKB was 24.7% higher than that of the wild strain after fermentation for 84 h,which would be beneficial for its application.In conclusion,the application of LB cloning system were verified using Sphingomonas sp.WG.The LB cloning system may provide an efficient tool for fusion expression of target genes.

Abstract:Gas vesicles are a unique class of gas-filled protein nanostructures which are commonly found in cyanobacteria and Halobacterium.The gas vesicles may scatter sound waves and generate harmonic signals,which enabled them to have the potential to become a novel ultrasound contrast agent.However,the current hypertonic cracking method for isolating gas vesicles contains tedious operational procedures and is of low yield,thus not suitable for large-scale application.To overcome these technical challenges,we developed a rapid and efficient method for isolating gas vesicles from Microcystis. The new H₂O₂-based method increased the yield by three times and shortened the operation time from 24 hours to 7 hours.The H₂O₂method is not only suitable for isolation of gas vesicles from laboratory-cultured Microcystis,but also suitable for colonial Microcystis covered with gelatinous sheath. The gas vesicles isolated by H₂O₂ method showed good performance in ultrasound contrast imaging.In conclusion,this new method shows great potential for large-scale application due to its high efficiency and wide adaptability,and provides technical support for developing gas vesicles into a biosynthetic ultrasonic contrast agent.

Abstract:Enzyme separation,purification,immobilization,and catalytic performance improvement have been the research hotspots and frontiers as well as the challenges in the field of biocatalysis.Thus,the development of novel methods for enzyme purification,immobilization,and improvement of their catalytic performance and storage are of great significance.Herein,ferritin was fused with the lichenase gene to achieve the purpose.The results showed that the fused gene was highly expressed in the cells of host strains,and that the resulted fusion proteins could self-aggregate into carrier-free active immobilized enzymes in vivo.Through low-speed centrifugation,the purity of the enzymes was up to>90%,and the activity recovery was 61.1%.The activity of the enzymes after storage for 608 h was higher than the initial activity.After being used for 10 cycles,it still maintained 50.0% of the original activity.The insoluble active lichenase aggregates could spontaneously dissolve back into the buffer and formed the soluble polymeric lichenases with the diameter of about 12 nm.The specific activity of them was 12.09 times that of the free lichenase,while the catalytic efficiency was 7.11 times and the half-life at 50℃ was improved 11.09 folds.The results prove that the ferritin can be a versatile tag to trigger target enzyme self-aggregation and oligomerization in vivo,which can simplify the preparation of the target enzymes,improve their catalysis performance,and facilitate their storage.

Abstract:Bio-separation engineering"is a compulsory course for undergraduate students majored in bioengineering,and an important part of the"emerging engineering education "system for bioengineering.Our teaching team follows the principle of" student development as the center,innovation thinking as the core".Guided by the concept of "learning achievement" ,we reconstructed the teaching contents of this course,and carried out the teaching reform aiming at solving several long-standing problems.These include,for instance,the theoretical teaching is separated from the experimental practice,and students cannot internalize the theoretical knowledge into practical ability in time.Moreover,the contents of course is out-of-date and out of line with industry demand,the teaching form and assessment methods are relatively single,and the students'professional ability and quality are not effectively cultivated.In the new curriculum system,in which the"online"and "offline" teaching are both applied,we broke the boundary between theoretical and experimental courses,and made the contents keep up with the forefront of industry development through research-based teaching.In terms of teaching methods and teaching evaluation,we made full use of modern information technology to enrich classroom teaching activities,and carried out complete,dynamic and diversified assessment for students.These teaching reform measures greatly improved the students'interest in learning this course,as well as their professionalism and research ability.

Abstract:Synthetic Biology is one of the most promising fields of modern Biology and a frontier interdisciplinary subject in the 21st century.With the rapid development of synthetic biology,the International Genetically Engineered Machine (iGEM) competition has emerged.The iGEM competition,based on the subject foundation of Synthetic Biology,intends to solve the biological problems in our daily life by applying modern biological technology.In recent years,with the continuous increase of participating teams,the iGEM competition has received extensive attention and achieved great progress.On the basis of the development of Synthetic Biology,we analyzed the 2018-2020 award-winning projects of the iGEM competition and illustrated the role and significance of the iGEM competition in cultivating college students'innovative thinking and ability with the participation experience of the iGEM team of Southwest Jiaotong University as an example.

Abstract:As an emerging branch of biology,Synthetic Biology has seen rapid development with great potential in theoretical research and application.With a lot of brand-new concepts and research methods,it brings challenges to university teachers,and little experience is available in China on the teaching of Synthetic Biology.In this study,we discussed the general education-based development and application of the course on Synthetic Biology (a discipline in "liberal arts" in Zhejiang University) from the background,design,implementation,outcome,and problems of the course,hoping to provide a reference for the optimization of the course and the design of similar courses in other universities in China.

Abstract:Teaching quality is directly related to the performance of universities in fostering talents.Being innovative,high-level,and challenging (IHC) is the basic goal of course reform at universities in the new era.It is essential to reform the contents and teaching mode to improve the IHC properties of the existing courses.We first designed the three-dimensional goals of Molecular Biology Experiment teaching and the contents to support these goals.Then,we pinpointed the common points shared by blended teaching and experiment course,and designed the ways of blended teaching for the course.The reformed course contents and teaching mode have enhanced its IHC properties,and achieved good teaching performance.This paper provides a reference for the reform of experiment courses in universities.

Abstract:As the content of static biochemistry is boring and traditional cramming education fails to draw the attention of students,we divided the course into small units and then made full use of "micro-teaching" resources such as multimedia slides,pictures,objects,experimental demonstrations and animations of the biochemical principles.The method created a"micro-teaching "environment,which combined boring static knowledge with vivid dynamic elements,thus producing a strong sensory impact on students,highlighting the knowledge in their mind,prompting students'participation,and greatly improving the teaching effects.This article introduces the goals and development process of this teaching method,and takes" Protein Denaturation and Renaturation "as an example to demonstrate the design and implementation process of the" micro-teaching",which seems to be of great significance to the improvement of the teaching quality of biochemistry course and serves as a reference for reform in other courses.

Abstract:General Biology is basic discipline in Environmental Science and Engineering,which lays a basis for the specialized courses in this major.As the course involves a wide range of knowledge,traditional "cramming education" cannot attract the interest of students.To meet the needs of talents against the backdrop of "double world-class project",our teaching team applied the"double-cycle teaching mode"of blended teaching in the first class along with ideological education,professional education,innovation and entrepreneurship education in the second class of this course.With this method,major headway has been made in this course.This study is expected to serve as a reference for reform in other relevant courses.

Abstract:Bioengineering majors require students to acquire excellent abilities of thinking and analyzing complex problems and have high requirements for students'comprehensive practical skills.Because of the professional characteristics,it is necessary to develop students'abilities to solve complex problems via the teaching of a series of experiments.Therefore,it is particularly important to reform the traditional experiment teaching for students majoring in bioengineering to improve the teaching quality,which have great significance for the cultivation of comprehensive talents.In this study,with the advantages of geographical location and resources to cultivate application-oriented innovative talents,the course group of Comprehensive Experiment of Bioengineering has designed the course based on virtual simulation technology in Binzhou University.Taking the experiment of extraction and bioactivity analysis of Suaeda salsa(growing in the Yellow River Delta) polysaccharide in fermentation as a case,we studied the course design idea,experimental process,teaching method and result analysis,and have improved the teaching performance.This case analysis provides new ideas and content reference for the teaching reform of similar courses.