Sheng wu gong cheng xue bao = Chinese journal of biotechnology最新文献

The WRKY transcription factor gene family is a plant-specific transcription factor that plays important roles defense responses. Studies in model plant Arabidopsis demonstrated that WRKYs function downstream of mitogen activated-protein kinase (MAPK) signaling cascade and participate in defense responses through activating the expression of defense-related genes. However, the roles of WRKYs in defense responses have not been previously investigated in paleopolyploidy soybean. Bioinfomatic analysis revealed that there are three pair of GmWRKY33 genes in the soybean genome. The identity of first two pair of GmWRKY33 genes is greater than 84% (named as GmWRKY33A). The identity of genes within the same pair is greater than 95%. A 300 bp fragment highly homologous to these four GmWRKY33A was chosen to clone into bean pod mosaic virus (BPMV)-based silencing vector (BPMV-VIGS) to achieve the goal of silencing four GmWRKY33A genes simultaneously. In this study, we simultaneously silenced four homologous genes of GmWRKY33A using a bean pod mottle virus (BPMV) vector carrying a single fragment of GmWRKY33A. Comparing the silenced plants with the vector control plants, no evident morphological phenotypes were observed. However, the GmWRKY33A-silenced plants exhibited significantly reduced resistance to Pseudomonas syringae pv. glycinea (Psg), Xanthomonas axonopodis pv. glycine (Xag), as well as to soybean mosaic virus (SMV). Furthermore, we demonstrated that silencing these GmWRKY33A genes significantly inhibited the activation of GmMPK3/GmMPK6 induced by Psg infection. Collectively, our results suggest that GmWRKY33As are involved in soybean immunity through regulating the transcription of GmMPK3/6 genes or activating the kinase activities of GmMPK3/6. Taken together, our results demonstrated that GmWRKY33As are positive regulators of soybean immune responses.

To explore the role and metabolic differences of plant hormones in regulating the flower bud size of daylily, we collected the flower buds from two daylily varieties 'Datong Huanghua' and 'Dongbei Huanghua' at the young bud, green, yellowing, and yellow stages for transcriptome sequencing. The differentially expressed genes (DEGs) were screened, and the exogenous plant hormone spraying experiments were conducted. A total of 199 DEGs related to the biosynthesis and metabolism of plant hormones was screened out at different flower development stages of 'Datong Huanghua' and 'Dongbei Huanghua'. These genes regulated the biosynthesis and metabolism of six plant hormones: abscisic acid, gibberellin, auxin, jasmonate, cytokinin, and ethylene. The DEGs associated with auxin were the most, which suggested that auxin played a role in regulating flower bud development. The auxin response factor (ARF) presented up-regulated expression at all the four stages of flower bud development, indicating that ARF played a positive regulatory role throughout the flower bud development of daylily. The experiments with exogenous spraying of six hormones further verified that indole-3-acetic acid (IAA) significantly promoted the growth and increased the nutrient content in the flower buds of 'Datong Huanghua', suggesting that IAA played a role in regulating flower bud development. Our results laid a theoretical foundation for probing into the regulatory mechanism of flower bud development of 'Datong Huanghua' and 'Dongbei Huanghua'.

Terpene synthases (TPSs) play a crucial role in the synthesis of terpenoids that contribute to the scent profiles of flowers. However, few studies report the genome-wide analysis of TPSs gene in Jasminum sambac var. Fuzhou bifoliatum and their expression pattern in response to methyl jasmonate (MeJA). In this study, we employed bioinformatics tools for genome-wide analysis of the J. sambac TPS (DJTPS) gene family and determined the physical and chemical properties, subcellular location, protein-protein interactions, phylogenetic relationship, subfamily classification, chromosomal location and collinearity, gene structure, conserved motifs, and promoter cis-acting elements. The expression patterns of DJTPSs in different tissues and in response to MeJA treatment were analyzed based on the transcriptome data combined with quantitative real-time PCR (qRT-PCR). We identified 32 intact DJTPS genes in the genome of J. sambac, which presented uneven distribution across nine chromosomes. All the deduced proteins were hydrophilic, predominantly localized in the cytoplasm. The phylogenetic analysis classified the DJTPS genes into five subfamilies: TPS-a, TPS-b, TPS-c, TPS-e/f, and TPS-g. The results of the collinearity analysis showed a total of 10 sets of replication events in DJTPSs, most of which underwent purifying selection. A comparative analysis of TPS homologous gene pairs was performed among J. sambac var. Fuzhou bifoliatum and other six species, which revealed different number of homologous gene pairs. The number of exons and motifs was conserved within the same subfamily. DJTPS genes carried multiple elements that may be involved in the response to MeJA. In addition, the transcriptome and qRT-PCR data unveiled that several TPS genes exhibited tissue-specific expression patterns, and the genes with specific expression in flowers were the most. Upon exposure to MeJA, 14 TPS genes showcased upregulated expression 5 h or 6 h post-treatment, and DJTPS03, DJTPS04 and DJTPS21 showed significantly increased expression levels after MeJA treatment. This study provides preliminary evidence that MeJA possesses the ability to enhance the expression of DJTPS genes during the critical flowering stage, which will facilitate the synthesis of terpenoids and improve the quality of floral fragrance.

Polyethylene terephthalate (PET) is one of the widely used plastics, but its waste pollution has become a global environmental issue. The discovery of polyethylene terephthalate hydrolase (PETase) has provided a green and environmentally friendly approach for PET degradation. However, PETase produces intermediate products that inhibit the enzyme's further activity, leading to a decrease in enzyme efficiency. Mono(2-hydroxyethyl) terephthalate hydrolase (MHETase) works synergistically with PETase to further degrade the intermediate product MHET into ethylene glycol (EG) and terephthalic acid (TPA). MHETase exhibits extremely high specificity for MHET and is crucial for the complete degradation of PET. This article comprehensively reviews MHETase from various perspectives, including its three-dimensional structure, substrate binding, and catalytic mechanism. It demonstrates the structural features and key residues associated with the enzyme's degrading activity and discusses the progress in enzyme engineering modifications. Additionally, the study envisions the development of a two-enzyme PET degradation system by combining MHETase with PETase, aiming to provide valuable references for designing and developing more efficient PET hydrolytic enzyme systems.

Tyrosinase is a copper-containing polyphenol oxidase widely applied in the food, cosmetics, pharmaceutical, and other industries. Currently, the production of commercial tyrosinase primarily relies on extraction from fungi, which has high costs, low purity, low specific activity, and poor stability. The objective of this study is to obtain highly expressed bacterial tyrosinase with potential for industrial applications. The bacterial tyrosinases from five different sources were heterologously expressed in Escherichia coli BL21(DE3), and the tyrosinases TyrBm and TyrVs derived from Bacillus megaterium and Verrucomicrobium spinosum were obtained with the enzyme activities of (16.1±0.2) U/mL and (48.6±0.9) U/mL, respectively. After protein purification, we compared the enzymatic properties of TyrBm and TyrVs, which revealed that TyrVs exhibited better thermal stability and higher substrate specificity than TyrBm. On the basis of characterizing TyrVs with high catalytic performance, we established a biological hair dyeing system based on TyrVs catalysis to achieve in-situ catalytic hair dyeing. The color washing fastness test measured the ∆E value less than 7.38±0.64 after simulated 14-day cleaning. To facilitate the rapid separation of catalytic products and enzymes, we successfully constructed an immobilized enzyme TyrVs-CipA dependent on self-assembly label CipA and applied this enzyme in the DOPA modification of hydrolyzed silk fibroin (HSF). The immobilized enzyme continuously catalyzed HSF for more than seven cycles, resulting in a single DOPA modification degree exceeding 70.00%. Further investigations demonstrated that DOPA modification enhances the scavenging activity of HSF towards DPPH and O₂^- radicals by 507.80% and 78.23%, respectively. This study provides a technical foundation for the development of environmentally friendly biological hair dye based on tyrosinase and biomaterials for tissue engineering.

The major science and technology infrastructure in the field of life science is an indispensable and important content in the large-science facility landscape. It encompasses cutting-edge, strategic, and fundamental aspects. This field differs from traditional facilities such as particle physics, astronomy and nuclear energy. Moreover, it represents a relatively underdeveloped area in China's facility landscape. Unique characteristics are observed in terms of capital investment, physical form, facility lifespan, digitization degree, organizational structure, project risk, and development effect when compared to traditional facilities. Despite its importance, challenges persist in project establishment, investment, management, and construction. Therefore, it is necessary to strengthen the condensing mechanism for addressing major scientific issues in the life science field, improve the strategic investment layout, facilitate the localization of technical equipment based on original scientific ideas, and strengthen the differentiated management capacity of life science facilities.

In recent years, artificial intelligence has been employed to empower synthetic biology, demonstrating great potential in the simulation and prediction of protein structures as well as the design and optimization of regulatory elements and metabolic networks. Integrating artificial intelligence into the teaching of Synthetic Biology is in line with the development trends of synthetic biology and can promote the cultivation of interdisciplinary high-level talents and collaborative innovation. This paper expounds the idea of integrating artificial intelligence into the teaching of Synthetic Biology from establishing interdisciplinary course contents and teaching methods, simultaneously considering the fundamentals and application of artificial intelligence in synthetic biology, cultivating independent learning and innovative practice abilities, and enhancing the ethics education related to artificial intelligence. Furthermore, a system integrating artificial intelligence with the teaching contents of Synthetic Biology is designed, which focuses on supplementing fundamentals of artificial intelligence and incorporating artificial intelligence into the classroom and experimental teaching contents of Synthetic Biology. Moreover, with the course of Synthetic Biology in Jiangnan University as an example, this paper presents the pathway of integrating artificial intelligence into the teaching of this course under the background of discipline crossing. Finally, the teaching effects are expected.

Under the background of developing new engineering disciplines, teaching reform is a key strategy applied by higher education institutions to develop new engineering professionals and accomplish the mission of cultivating morality and nurturing talents. As a foundational course for majors of life sciences and food sciences, "Principles of Fermentation Engineering" has a strong scientific, practical, and historical focus. It serves as an excellent resource for changing the way that college students are taught professional courses. To examine the reform and practical route of specialized course teaching combined with innovation and entrepreneurship fostering under the integration production, education, and research, this article takes the teaching of "Principles of Fermentation Engineering" for undergraduates majoring in food science and engineering at Hebei Agricultural University as an example. A new teaching paradigm integrating production, education, and research is developed considering a variety of factors, including instructional content, teaching methods, and evaluation approaches. This paradigm is result-oriented, replaces examination with competition, and promotes learning by research. It achieves the integration of specialized course teaching and innovation and entrepreneurship fostering and lays a foundation for the teaching reform and the development of professional talents in the context of developing new engineering disciplines.

Ganoderma lucidum is a precious fungus with both edible and medicinal values and has a long history of medical use. Triterpenes as the main active components endow G. lucidum with anti-tumor, antioxidant, and other pharmacological activities. The present study endeavors to establish a proficient liquid-state fermentation technology for the enhanced production of triterpenes. In view of the limitations inherent in conventional submerged fermentation and oscillation-static two-stage cultivation, this study established an oscillation-static cycle cultivation process and optimized the cultivation conditions by building an artificial neural network model based on genetic algorithms. The cultivation conditions for the high-yield production of triterpenes were optimized as follows: 2.8 days of oscillation, 7.3 days of static cultivation, 0.2 day of oscillation, and 0.3 day of static cultivation. Under these conditions, the content of triterpenes reached 20.82 mg/g. The yield of triterpenes reached 129.09 mg/L, showing a remarkable increase of 324.78% compared with that of the Z10J0 method. Moreover, the established method shortened the cultivation cycle by 10.6 days. The mycelia cultivated under this regimen exhibited commendable anti-tumor and antioxidant activities. This study not only presents an economical liquid-state fermentation approach but also streamlines the fermentation flow, reduces fermentation duration, and effectively ameliorates drawbacks associated with conventional cultivation methods. In addition, this study gives valuable insights into the scaled application of liquid-state fermentation in the high-yield production of triterpenes, which showcases broad prospects.

Value shaping and innovation capability improvement are two important goals of postgraduate course teaching. Biocatalysis and Enzyme Engineering is a core course for postgraduates majoring in bioengineering. Our teaching team established a "dual-drive and dual-guide" teaching model, that is, "double drive" (value-driven + innovation-driven) as the traction, this model integrated professional quality with industry needs, engineering ethics, and scientist spirit, combined theoretical teaching with research frontiers, industrial cases, and engineering practice, and incorporated thematic lectures, group peer evaluation, and review papers into course assessment. The teaching under this model achieved the teaching objectives of "double guide" (guiding ideology and ability), and improved the postgraduates' value identification, innovation awareness, engineering thinking, and ability to solve practical engineering problems.