1.Identification and expression analysis of TCP family members in tobacco (Nicotiana tabacum L.).
Shize WANG ; Yun LI ; Yucui HAN ; Shizhou YU ; Shuang WANG ; Yong LIU ; Xiaohu LIN
Chinese Journal of Biotechnology 2024;40(1):226-238
		                        		
		                        			
		                        			TCP family as plant specific transcription factor, plays an important role in different aspects of plant development. In order to screen TCP family members in tobacco, the homologous sequences of tobacco and Arabidopsis TCP family were identified by genome-wide homologous alignment. The physicochemical properties, phylogenetic relationships and cis-acting elements were analyzed by bioinformatics. The homologous genes of AtTCP3/AtTCP4 were screened, and RT-qPCR was used to detect the changes of gene expression upon 20% PEG6000 treatment. The results show that tobacco contains 63 TCP family members. Their amino acid sequence length ranged from 89 aa to 596 aa, and their protein hydropathicity grand average of hydropathicity (GRAVY) ranged from -1.147 to 0.125. The isoelectric point (pI) ranges from 4.42 to 9.94, the number of introns is 0 to 3, and the subcellular location is all located in the nucleus. The results of conserved domain and phylogenetic relationship analysis showed that the tobacco TCP family can be divided into PCF, CIN and CYC/TB1 subfamilies, and each subfamily has a stable sequence. The results of cis-acting elements in gene promoter region showed that TCP family genes contain low docile acting elements (LTR) and a variety of stress and metabolic regulation related elements (MYB, MYC). Analysis of gene expression patterns showed that AtTCP3/AtTCP4 homologous genes (NtTCP6, NtTCP28, NtTCP30, NtTCP33, NtTCP42, NtTCP57, NtTCP63) accounted for 20% PEG6000 treatment significantly up-regulated/down-regulated expression, and NtTCP30 and NtTCP57 genes were selected as candidate genes in response to drought. The results of this study analyzed the TCP family in the tobacco genome and provided candidate genes for the study of drought-resistance gene function and variety breeding in tobacco.
		                        		
		                        		
		                        		
		                        			Nicotiana/genetics*
		                        			;
		                        		
		                        			Phylogeny
		                        			;
		                        		
		                        			Plant Breeding
		                        			;
		                        		
		                        			Amino Acid Sequence
		                        			;
		                        		
		                        			Arabidopsis
		                        			;
		                        		
		                        			Polyethylene Glycols
		                        			
		                        		
		                        	
2.Commentary: polymer binding modules accelerate enzymatic degradation of poly(ethylene terephthalate).
Yi LU ; Ruizhi HAN ; Ulrich SCHWANEBERG ; Yu JI
Chinese Journal of Biotechnology 2023;39(5):1883-1888
		                        		
		                        			
		                        			The large scale production and indiscriminate use of plastics led to serious environmental pollution. To reduce the negative effects of plastics waste on the environment, an approach of enzymatic degradation was put forward to catalyze plastics degradation. Protein engineering strategies have been applied to improve the plastics degrading enzyme properties such as activity and thermal stability. In addition, polymer binding modules were found to accelerate the enzymatic degradation of plastics. In this article, we introduced a recent work published in Chem Catalysis, which studied the role of binding modules in enzymatic hydrolysis of poly(ethylene terephthalate) (PET) at high-solids loadings. Graham et al. found that binding modules accelerated PET enzymatic degradation at low PET loading (< 10 wt%) and the enhanced degradation cannot be observed at high PET loading (10 wt%-20 wt%). This work is beneficial for the industrial application of polymer binding modules in plastics degradation.
		                        		
		                        		
		                        		
		                        			Polyethylene Terephthalates/metabolism*
		                        			;
		                        		
		                        			Polymers
		                        			;
		                        		
		                        			Plastics
		                        			;
		                        		
		                        			Ethylenes
		                        			
		                        		
		                        	
3.Polyethylene biodegradation: current status and perspectives.
Liting ZHANG ; Bo ZHANG ; Weidong XU ; Zhongli CUI ; Hui CAO
Chinese Journal of Biotechnology 2023;39(5):1949-1962
		                        		
		                        			
		                        			Polyethylene (PE) is the most abundantly used synthetic resin and one of the most resistant to degradation, and its massive accumulation in the environment has caused serious pollution. Traditional landfill, composting and incineration technologies can hardly meet the requirements of environmental protection. Biodegradation is an eco-friendly, low-cost and promising method to solve the plastic pollution problem. This review summarizes the chemical structure of PE, the species of PE degrading microorganisms, degrading enzymes and metabolic pathways. Future research is suggested to focus on the screening of high-efficiency PE degrading strains, the construction of synthetic microbial consortia, the screening and modification of degrading enzymes, so as to provide selectable pathways and theoretical references for PE biodegradation research.
		                        		
		                        		
		                        		
		                        			Polyethylene/metabolism*
		                        			;
		                        		
		                        			Bacteria/metabolism*
		                        			;
		                        		
		                        			Plastics/metabolism*
		                        			;
		                        		
		                        			Biodegradation, Environmental
		                        			;
		                        		
		                        			Microbial Consortia
		                        			
		                        		
		                        	
4.Advances in poly(ethylene terephthalate) hydrolases.
Zhiyi ZHAO ; Guoqiang ZHANG ; Kun LIU ; Shengying LI
Chinese Journal of Biotechnology 2023;39(5):1998-2014
		                        		
		                        			
		                        			Plastics have brought invaluable convenience to human life since it was firstly synthesized in the last century. However, the stable polymer structure of plastics led to the continuous accumulation of plastic wastes, which poses serious threats to the ecological environment and human health. Poly(ethylene terephthalate) (PET) is the most widely produced polyester plastics. Recent researches on PET hydrolases have shown great potential of enzymatic degradation and recycling of plastics. Meanwhile, the biodegradation pathway of PET has become a reference model for the biodegradation of other plastics. This review summarizes the sources of PET hydrolases and their degradation capacity, degradation mechanism of PET by the most representative PET hydrolase-IsPETase, and recently reported highly efficient degrading enzymes through enzyme engineering. The advances of PET hydrolases may facilitate the research on the degradation mechanism of PET and further exploration and engineering of efficient PET degradation enzymes.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Hydrolases/metabolism*
		                        			;
		                        		
		                        			Polyethylene Terephthalates/metabolism*
		                        			;
		                        		
		                        			Plastics/metabolism*
		                        			;
		                        		
		                        			Ethylenes
		                        			
		                        		
		                        	
5.Expression, purification and characterization of a novel bis (hydroxyethyl) terephthalate hydrolase from Hydrogenobacter thermophilus.
Yangyang CHEN ; Jian GAO ; Yipei ZHAO ; Hao WANG ; Xu HAN ; Jie ZHANG ; Qun GU ; Ying HOU ; Weidong LIU
Chinese Journal of Biotechnology 2023;39(5):2015-2026
		                        		
		                        			
		                        			PET (polyethylene terephthalate) is one of the most important petrochemicals that is widely used in mineral water bottles, food and beverage packaging and textile industry. Because of its stability under environmental conditions, the massive amount of PET wastes caused serious environmental pollution. The use of enzymes to depolymerize PET wastes and upcycling is one of the important directions for plastics pollution control, among which the key is the depolymerization efficiency of PET by PET hydrolase. BHET (bis(hydroxyethyl) terephthalate) is the main intermediate of PET hydrolysis, its accumulation can hinder the degradation efficiency of PET hydrolase significantly, and the synergistic use of PET hydrolase and BHET hydrolase can improve the PET hydrolysis efficiency. In this study, a dienolactone hydrolase from Hydrogenobacter thermophilus which can degrade BHET (HtBHETase) was identified. After heterologous expression in Escherichia coli and purification, the enzymatic properties of HtBHETase were studied. HtBHETase shows higher catalytic activity towards esters with short carbon chains such as p-nitrophenol acetate. The optimal pH and temperature of the reaction with BHET were 5.0 and 55 ℃, respectively. HtBHETase exhibited excellent thermostability, and retained over 80% residual activity after treatment at 80 ℃ for 1 hour. These results indicate that HtBHETase has potential in biological PET depolymerization, which may facilitate the enzymatic degradation of PET.
		                        		
		                        		
		                        		
		                        			Hydrolases/metabolism*
		                        			;
		                        		
		                        			Bacteria/metabolism*
		                        			;
		                        		
		                        			Hydrolysis
		                        			;
		                        		
		                        			Polyethylene Terephthalates/metabolism*
		                        			
		                        		
		                        	
6.Enzymatic properties and degradation characterization of a bis(2-hydroxyethyl) terephthalate hydrolase from Saccharothrix sp.
Jie ZHANG ; Ruida SHAN ; Xia LI ; Zhixiong ZENG ; Dengyue SUN
Chinese Journal of Biotechnology 2023;39(5):2027-2039
		                        		
		                        			
		                        			The discovery of new enzymes for poly(ethylene terephthalate) (PET) degradation has been a hot topic of research globally. Bis-(2-hydroxyethyl) terephthalate (BHET) is an intermediate compound in the degradation of PET and competes with PET for the substrate binding site of the PET-degrading enzyme, thereby inhibiting further degradation of PET. Discovery of new BHET degradation enzymes may contribute to improving the degradation efficiency of PET. In this paper, we discovered a hydrolase gene sle (ID: CP064192.1, 5085270-5086049) from Saccharothrix luteola, which can hydrolyze BHET into mono-(2-hydroxyethyl) terephthalate (MHET) and terephthalic acid (TPA). BHET hydrolase (Sle) was heterologously expressed in Escherichia coli using a recombinant plasmid, and the highest protein expression was achieved at a final concentration of 0.4 mmol/L of isopropyl-β-d-thiogalactoside (IPTG), an induction duration of 12 h and an induction temperature of 20 ℃. The recombinant Sle was purified by nickel affinity chromatography, anion exchange chromatography, and gel filtration chromatography, and its enzymatic properties were also characterized. The optimum temperature and pH of Sle were 35 ℃ and 8.0, and more than 80% of the enzyme activity could be maintained in the range of 25-35 ℃ and pH 7.0-9.0 and Co2+ could improve the enzyme activity. Sle belongs to the dienelactone hydrolase (DLH) superfamily and possesses the typical catalytic triad of the family, and the predicted catalytic sites are S129, D175, and H207. Finally, the enzyme was identified as a BHET degrading enzyme by high performance liquid chromatography (HPLC). This study provides a new enzyme resource for the efficient enzymatic degradation of PET plastics.
		                        		
		                        		
		                        		
		                        			Actinomycetales/genetics*
		                        			;
		                        		
		                        			Hydrolases/metabolism*
		                        			;
		                        		
		                        			Phthalic Acids/chemistry*
		                        			;
		                        		
		                        			Polyethylene Terephthalates/metabolism*
		                        			
		                        		
		                        	
7.Engineering the plastic degradation enzyme Ple629 from marine consortium to improve its thermal stability.
Yipei ZHAO ; Hao WANG ; Pan WU ; Zhishuai LI ; Fufeng LIU ; Qun GU ; Weidong LIU ; Jian GAO ; Xu HAN
Chinese Journal of Biotechnology 2023;39(5):2040-2052
		                        		
		                        			
		                        			Petrochemical-derived polyester plastics such as polyethylene terephthalate (PET) and polybutylene adipate terephthalate (PBAT) have been widely used. However, the difficulty to be degraded in nature (PET) or the long biodegradation cycle (PBAT) resulted in serious environmental pollution. In this connection, treating these plastic wastes properly becomes one of the challenges of environment protection. From the perspective of circular economy, biologically depolymerizing the waste of polyester plastics and reusing the depolymerized products is one of the most promising directions. Recent years have seen many reports on polyester plastics degrading organisms and enzymes. Highly efficient degrading enzymes, especially those with better thermal stability, will be conducive to their application. The mesophilic plastic-degrading enzyme Ple629 from the marine microbial metagenome is capable of degrading PET and PBAT at room temperature, but it cannot tolerate high temperature, which hampers its potential application. On the basis of the three-dimensional structure of Ple629 obtained from our previous study, we identified some sites which might be important for its thermal stability by structural comparison and mutation energy analysis. We carried out transformation design, and performed expression, purification and thermal stability determination of the mutants. The melting temperature (Tm) values of mutants V80C and D226C/S281C were increased by 5.2 ℃ and 6.9 ℃, respectively, and the activity of mutant D226C/S281C was also increased by 1.5 times compared with that of the wild-type enzyme. These results provide useful information for future engineering and application of Ple629 in polyester plastic degradation.
		                        		
		                        		
		                        		
		                        			Plastics/metabolism*
		                        			;
		                        		
		                        			Polyethylene Terephthalates/metabolism*
		                        			;
		                        		
		                        			Biodegradation, Environmental
		                        			;
		                        		
		                        			Metagenome
		                        			
		                        		
		                        	
8.Research Advances in Medical Materials and Products for Soft Tissue Repairs.
Jiaqi LI ; Rui WANG ; Qianqian HAN ; Xue SUN
Chinese Journal of Medical Instrumentation 2023;47(4):415-423
		                        		
		                        			
		                        			Soft tissue is an indispensable tissue in human body. It plays an important role in protecting the body from external physical, chemical or biological factors. Mild soft tissue injuries can self-heal, while severe soft tissue injuries may require related treatment. Natural polymers (such as chitosan, hyaluronic acid, and collagen) and synthetic polymers (such as polyethylene glycol and polylactic acid) exhibit good biocompatibility, biodegradability and low toxicity. It can be used for soft tissue repairs for antibacterial, hemostatic and wound healing purposes. Their related properties can be enhanced through modification or preparation of composite materials. Commonly used soft tissue repairs include wound dressings, biological patches, medical tissue adhesives, and tissue engineering scaffolds. This study introduces the properties, mechanisms of action and applications of various soft tissue repair medical materials, including chitosan, hyaluronic acid, collagen, polyethylene glycol and polylactic acid, and provides an outlook on the application prospects of soft tissue repair medical materials and products.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Biocompatible Materials/chemistry*
		                        			;
		                        		
		                        			Chitosan/chemistry*
		                        			;
		                        		
		                        			Hyaluronic Acid
		                        			;
		                        		
		                        			Tissue Scaffolds/chemistry*
		                        			;
		                        		
		                        			Collagen/chemistry*
		                        			;
		                        		
		                        			Polymers/chemistry*
		                        			;
		                        		
		                        			Polyethylene Glycols
		                        			;
		                        		
		                        			Soft Tissue Injuries
		                        			
		                        		
		                        	
9.Meta-analysis of different joint interfaces in total hip arthroplasty under long-term follow-up.
Gai-Ge WU ; Ling-An HUANG ; Li GUO ; Peng-Cui LI ; Xiao-Chun WEI
China Journal of Orthopaedics and Traumatology 2023;36(2):165-171
		                        		
		                        			OBJECTIVE:
		                        			To compare the long-term follow-up effect and complications of ceramic on ceramic (CoC) interface and ceramic on polyethyleneon ceramic (CoP) interface in primary total hip arthroplasty, and provide clinical evidence.
		                        		
		                        			METHODS:
		                        			Search PubMed, EMBase, the CoChrane Library databases, Web of science, Wanfang database, and CNKI from January 2000 to September 2021, screening and inclusion of randomized controlled trials (RCTs) comparing the long-term efficacy and complications of CoC interface and CoP interface in total hip arthroplasty. Literature screening, quality evaluation and data extraction were carried out according to the inclusion and exclusion criteria, using Review Manager 5.3 statistical software. The software was used to perform statistical analysis on joint function, revision, prosthesis fracture, abnormal joint noise, and prosthesis wear rate after CoC or CoP.
		                        		
		                        			RESULTS:
		                        			Seven RCTs studies were included, including 390 cases of hips with CoC artificial joints and 384 cases of hips with CoP artificial joints. The long-term joint function improvement of CoC and CoP artificial joints was similar and there was no significant differences, with an average difference was MD=0.63, 95%CI=(-1.81, 3.07), P=0.61. About the postoperative complications, CoC artificial joints have higher incidence rate of abnormal joint noise, with odds ratio (OR)=11.05, 95%CI=(2.04, 59.84), P=0.005. CoP artificial joints wear faster, with an average MD=-87.11, 95%CI=(-114.40, -59.82), P<0.000 1. There was no significant difference between the two groups in the replacement-related complications such as joint dislocation, prosthesis loosening, osteolysis, and the rate of prosthesis revision caused by various reasons.
		                        		
		                        			CONCLUSION
		                        			The clinical function results and complications of CoC artificial joints are comparable to those of CoP artificial joints. Although CoP artificial joint prosthesis has a faster wear rate, it does not affect joint function and increase complications, and there is no abnormal joint noise. CoC is expensive and the long-term efficacy is equivalent to CoP. Clinicians should consider cost performance when choosing CoC.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Arthroplasty, Replacement, Hip/methods*
		                        			;
		                        		
		                        			Hip Prosthesis
		                        			;
		                        		
		                        			Follow-Up Studies
		                        			;
		                        		
		                        			Prosthesis Design
		                        			;
		                        		
		                        			Polyethylene
		                        			;
		                        		
		                        			Prosthesis Failure
		                        			;
		                        		
		                        			Reoperation
		                        			;
		                        		
		                        			Ceramics
		                        			;
		                        		
		                        			Treatment Outcome
		                        			
		                        		
		                        	
10.Establishment of a quantitative method for GC analysis of polyoxyethylene (35) castor oil in microemulsion extracts.
Yan-Jing WANG ; Yi CHENG ; Ze-Min OU ; Yao ZHANG ; Lin YAN ; Yan TONG ; Jin-Yu WANG ; De-Wen LIU
China Journal of Chinese Materia Medica 2023;48(22):6075-6081
		                        		
		                        			
		                        			With the continuous exploration of microemulsions as solvents for traditional Chinese medicine extraction, polyoxyethy-lene(35) castor oil(CrEL), a commonly used surfactant, is being utilized by researchers. However, the problem of detecting residues of this surfactant in microemulsion extracts has greatly hampered the further development of microemulsion solvents. Based on the chemical structures of the components in CrEL and the content determination method of castor oil in the 2020 edition of the Chinese Pharmacopoeia(Vol. Ⅳ), this study employed gas chromatography(GC) and single-factor experiments to optimize the preparation method of methyl ricinoleate from CrEL. The conversion coefficient between the two was validated, and the optimal sample preparation method was used to process microemulsion extracts of Zexie Decoction from three batches. The content of methyl ricinoleate generated was determined, and the content of CrEL in the microemulsion extracts of Zexie Decoction was calculated using the above conversion coefficient. The results showed that the optimal preparation method for CrEL was determined. Specifically, 10 mL of 1 mol·L~(-1) KOH-methanol solution was heated at 60 ℃ for 15 min in a water bath. Subsequently, 10 mL of boron trifluoride etherate-methanol(1∶3) solution was heated at 60 ℃ for 15 min in a water bath, followed by extraction with n-hexane twice. CrEL could stably produce 20.84% methyl ricinoleate. According to this conversion coefficient, the average mass concentration of CrEL in the three batches of Zexie Decoction microemulsion extracts was 11.94 mg·mL~(-1), which was not significantly different from the CrEL mass concentration of 11.57 mg·mL~(-1) during microemulsion formulation, indicating that the established content determination method of this study was highly accurate, sensitive, and repeatable. It can be used for subsequent research on microemulsion extracts of Zexie Decoction and provide a reference for quality control of other drug formulations containing CrEL.
		                        		
		                        		
		                        		
		                        			Polyethylene Glycols/chemistry*
		                        			;
		                        		
		                        			Castor Oil
		                        			;
		                        		
		                        			Methanol
		                        			;
		                        		
		                        			Surface-Active Agents/chemistry*
		                        			;
		                        		
		                        			Solvents
		                        			;
		                        		
		                        			Water/chemistry*
		                        			;
		                        		
		                        			Emulsions/chemistry*
		                        			
		                        		
		                        	
            
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