Objective: Unfolded Protein Response (UPR) is one of the most important protein quality control mechanisms, the information about UPR with help to optimize heterologous protein synthesis by secretory pathway and cell self protection from inhibitory substances such as acid and ethanol.Methods: The indicating strains, W303- 1A (leu 2::UPRE-lac Z) and An-a (leu 2::UPRE-lac Z), abbreviated as WZ and AZ, respectively, were first constructed by CRISPR/Cas9 technology and using UPRE-lac Z as a reporter.Results: The growth curve test showed that there was no significant difference between two strains and their host ones. The β-galactosidase activity was assayed at 4 h of incubation time after adding four reagents into media as followings, 1 μg/ml tunicamycin, 8%(v/v) ethanol, 0.3%(v/v) acetic acid, 5% (v/v) ethanol + 0.1%(v/v)acetic acid, respectively. The values of strain AZ were 8.2, 26.4, 1.1 and 7.9 times of that of blank control, respectively, while the results of strain WZ were 12.6, 2.4, 1.0 and 1.0 times, respectively. Further β-glucosidase was expressed by YEplac195 vector into strain AZ and WZ cell. The assay results after 24 h cultivation in 2% cellobiose media showed that the β-glucosidase activities were 0.35 and 6.12 U/ml, respectively, and the responding LacZ values were 3.1 and 5.4 times of that of blank control, respectively.Conclusions: There exists significant difference on the UPR response spectrum between two strains, which also implies their divergent direction and strategy of genetic modification. This study lays the foundation of analytical methods for investigating the key limiting factors in cell inhibitor tolerance and heterologous protein expression in order to make better use of yeast ER and UPR pathway engineering.
Background: An eukaryotic cell relies on its cellular organelles and organelles derived compartments to perform complicated biochemical reactions efficiently. One valid way to reveal the efficient cooperation between each organelle is to isolate distinct organelles. Although several techniques are developed to separate organelles, there is almost no easy method to assess the isolation process. Goal: Construct a strain of Saccharomyces cerevisiae to evaluate the efficiency of subcellular fractionation. Methods: A detection strain of Saccharomyces cerevisiae was constructed by traditional molecular biological and cell biological methods. One soluble protein and ten membrane proteins, chosen to represent subcellular compartments as well as plasma membrane, were grouped and labeled with different epitope tags in one strain. Immunofluorescence results were compared with fluorescent protein fusions to evaluate the impact of epitope tags on the localization of these proteins. Finally, density gradient centrifugation was used to illustrate the usability of the detection strain. Results: The detetion strain labeling all major subcellular compartments of Saccharomyces cerevisiae was constructed successfully. All epitope tagged organelle markers localized to the intended subcellular sites. Each compartment of Saccharomyces cerevisiae could be detected after density gradient centrifugation. Conclusion: The detection strain is a convenient tool for the evaluation of subcellular fractionation results. It is potentially useful for future research of Saccharomyces cerevisiae cell biology.
Infectious disease mainly refer to transmissible diseases caused by pathogenic microorganisms. It can affect human health and even cause serious social crisis. In recent years, the outbreaks of infectious diseases like COVID-19 and ebola have prompted people to seek more efficient and convenient means to control and prevent infectious diseases. As an effective way currently, antibody has drawn people’s attention. However, palivizumab, for the prevention and treatment of respiratory syncytial virus, is the only one monoclonal antibody approved for infectious diseases. Nano-antibody (Nb) is the smallest known functional antibody capable of stably binding to antigens. Nb has the advantages of high stability, strong hydrophilicity, easy production through microbial systems, easy modification, etc. Due to its unique molecular properties, Nb has shown promising application prospects in the prevention, diagnosis and treatment of infectious diseases caused by viruses, bacteria, parasites, etc. Related studies have shown that Nb has good therapeutic effects on AIDS, influenza, novel coronavirus, etc. This paper focuses on the structural characteristics of Nb and its research progress in infectious diseases.
Recently, antibody drugs have developed rapidly in the field of biopharmaceuticals. Clinically, the type and number of monoclonal antibody-based tumor therapies are increasing. With confirmed structure of PD-1/PD-L1 protein,monoclonal antibody drugs against immune checkpoint PD-1/PD-L1 have been continuously developed and applied to the treatment of many principal cancers with high mortality and low cure rate. However, due to the complex physical and chemical properties and serious homogenization, monoclonal antibody drugs are necessary to screen out stable protein preparations for clinical applications based on the characteristics of different monoclonal antibody drugs. This article reviews the role of different antibody drug formulations (buffer components, drug excipients), combined with the PD-L1 target, which introduces the stability development of antibody drug formulations and the main points for evaluation of CDE.
CRISPR/Cas9, a new gene editing technology, mainly modifies the genetic information of organisms at the DNA level and has powerful gene editing ability. Now, it has been widely used in many fields, including gene function research, animal model construction, new breed breeding and gene therapy. The continuous development of CRISPR/Cas9 technology has brought a revolutionary breakthrough in the field of biology and medicine. Using this technology to construct gene mutant mice is not only conducive to the research of gene function, but also has an important reference value for the treatment of genetic diseases. In addition, this technology can effectively improve the production performance of livestock at the molecular level, and improve the disease resistance of livestock. Mainly introduces the research process, structure and classification of CRISPR/Cas system, expounds the mechanism of CRISPR/Cas9 technology and its application in animal gene editing, discusses the problems and optimization strategies of CRISPR/Cas9 in making gene editing animals, and prospects the development of CRISPR/Cas9.
Silent information regulator the factor 1 SIRT1(SIRT1), is NAD+ dependent enzymes with deacetylase activity, which participates in the regulation of many physiological functions of cells through substrate deacetylation, playing important roles in the process of glycolipid metabolism, aging, apoptosis, oxidative stress and so on. In addition, some studies have shown that SIRT1 is an important factor in the regulation of ovarian aging, follicular development and oocyte maturation. Moreover, the decrease of SIRT1 expression or the change of SIRT1 activity will lead to the aging of oocytes and the reduction of animal fertility. Therefore, to fully understand the function of SIRT1, and delay the aging of ovary and oocyte by regulating SIRT1 activity, thereby improving animal fertility, this paper describes the activation of SIRT1 and its biological processes involved in intracellular regulation, and discusses the main functions of SIRT1 from the perspective of energy metabolism, antioxidant stress and chromatin remodeling, highlights the regulatory roles of SIRT1 in animal follicular development and oocyte maturation.
Inspired by the adhesion of mussels, the researchers found that dopamine self-polymerizes to form polydopamine under alkaline conditions. Due to its strong adhesion properties, polydopamine has been widely used to modify various biological materials in the past few years. Most of the materials used in nerve repair are polymers, but the effect of using polymers alone to repair nerves is not good. Polydopamine modified polymers have better hydrophilicity and biocompatibility than single polymers. In addition, the polydopamine coating on the polymer can be used to further modify the molecules that promote nerve repair. This article reviews the synthesis mechanism and performance of polydopamine and the research progress of polydopamine-modified polymers in nerve repair, and at the end of the article, the development prospects of such materials are prospected.
Methanol is an abundant raw material with low price and high reduction degree, which is expected to be a promising feedstock for the next generation biomanufacturing. To convert methanol into value-added chemicals, constructing recombinant microorganisms using synthetic biology has received broad attention in recent years. However, due to the complex regulation of methanol metabolism, the current engineered synthetic methylotrophy still cannot use methanol as the sole carbon source for growth and chemical production. Based on the analysis of methanol metabolic mechanism in natural methylotrophy, this review summarized the main challenges for designing and constructing synthetic methylotrophy and proposed potential strategies to overcome these barriers. Especially, it focused on the following aspects, including: screening and engineering methanol dehydrogenase; optimizing and balancing formaldehyde assimilation pathways; bioconversion of methanol to chemicals.
Phloretin and its glycosides are natural dihydrochalcones with various physiological activities, such as antioxidant, anti-inflammatory and anti-bacterial activities, and have potential application in food, pharmaceutical, and cosmetics industries. At present, phloretin and its glycosides are mainly extracted from plants. However, the low content and complex components make it difficult to prepare products with high efficiency and low cost. With the development of metabolic engineering and synthetic biology, microbial production of phloretin and its glycosides has become an alternative approach. This article reviews the identification of key genes, the reconstruction of synthetic pathways and optimization strategies for the biosynthesis of phloretin and its glycosides. Finally, the potential strategies to solve existing problems as unspecific enzymes and the formation of multiple byproducts were proposed.
Degraded from alginate, alginate Oligosaccharides (AOS) is widely applies into various fields due to its physiological activities such as antioxidation, antitumor, bacteriostatic effect, immune regulation, promotion of cell growth and so on. The main preparation methods include physical degradation, chemical degradation and biological degradation, and brief comparison of these methods is summarized in this review. This paper mainly concludes biological preparation of AOS, including enzymatic hydrolysis by alginate lyase, microbial whole-cell fermentation and biosynthesis. Genetic engineering is of great significance in constructing genetically modified strain to improve the efficiency of biological method, so it is introduced in this review. In addition, the case of medium scale AOS biological preparation is also scientifically cited in this paper, and the future research directions of industrial production of AOS is prospected briefly. Realizing recommendations above will provide reference material for the future industrial preparation and applications of AOS.
Industrial development is inseparable from the formulation and release of industrial policies. Industrial policies are oriented and supportive to the industry, and the main bodies of their formulation include the central government and local governments. Taking the biological industry as an example, we collected and sorted out the biological industry policies issued by the central government and five local governments in Hubei, Shenzhen, Chongqing, Shanghai, and Beijing. The method of quantitative analysis was used to analyze the differences in external attributes and characteristics. The 27 national-level bio-industry structural policies used content analysis methods to classify and analyze policy tools, and constructed a two-dimensional policy tool analysis framework of “Policy Tools-Bioindustry Development Stages” to explore the differences in the use of policy tools between different industrial development stapes. The study found that the central government and local governments have different years of intensive release of bio-industry policies, and they are related to key nodes in the development of the bio-industry; there is a large gap in the frequency of use of different types of policy tools in bio-industry policies at the national level. Enviromental policy tools are the main ones, and demand-based policy tools are rarely used. At the same time, in response to the research results, relevant policy recommendations are put forward based on the development of my country’s biological industry and regional differences.
The emergence and continuous evolution of drug-resistant bacteria, especially multi-drug resistant bacteria, poses a huge threat to human health. With the gradual loss of special effects of antibiotics, the scientific community and the medical community have turned their eyes to antibacterial natural organisms-bacteriophage, and in some studies have proved that bacteriophage can be used as a new weapon to replace antibiotics to treat drug-resistant bacterial infections. Through statistics and analysis of the world patent applications of phage therapy and derived lyase therapy, information on patent development trends, applicant distribution characteristics and main patent applicants was obtained, and the main patent technology routes and hotspots of bacteriophage and lyase therapy were analyzed in detail.
Alzheimer’s disease is a progressive neurodegenerative disease with occult onset. With the global aging becoming more and more serious, Alzheimer’s disease is a serious threat to people’s health and quality of life. The causes of Alzheimer’s disease are complex, and the treatment of Alzheimer’s disease is still an important problem in medicine. In this paper, with the literature of Alzheimer’s disease gene research indexed in the core collection of Web of Science, bibliometric analysis of publication trends, researchers, institutions, countries and research hotspots was carried out to provide important reference for researchers in related fields.
