1.Detection methods for polyethylene terephthalate degrading enzymes: a review.
Hanxiao ZHANG ; Yunjie XIAO ; Haitao YANG ; Zefang WANG
Chinese Journal of Biotechnology 2023;39(8):3219-3235
Polyethylene terephthalate (PET) is one of the most widely used synthetic polyester. It poses serious threat to terrestrial, aquatic ecosystems and human health since it is difficult to be broken down and deposited in the environment. The biodegradation based on enzymatic catalysis offers a sustainable method for recycling PET. A number of PET hydrolases have been discovered in the last 20 years, and protein engineering has increased their degradation capabilities. However, no PET hydrolases that are practical for widespread industrial use have been identified. Screening of PET hydrolase using conventional detection techniques is laborious and inefficient process. Effective detection techniques are required to promote the commercialization of PET hydrolases. Using efficient detection techniques to screen potent industrial enzymes is essential for supporting the widespread industrial implementation of PET hydrolases. To define PET hydrolase, scientists have created a number of analytical techniques recently. The detection techniques that can be used to screen PET hydrolase, including high performance liquid chromatography, ultraviolet absorption spectrometric, and fluorescence activated droplet sorting method, are summarized in this study along with their potential applications.
Humans
;
Polyethylene Terephthalates
;
Ecosystem
;
Biodegradation, Environmental
;
Catalysis
;
Hydrolases
2.Expression, thermal stability modification and application in PHB degradation of polyhydroxyalkanoate depolymerase from Thermomonospora umbrina.
Zhigang LI ; Shiheng CHEN ; Demin KONG ; Sheng CHEN ; Lei WANG ; Jing WU
Chinese Journal of Biotechnology 2023;39(8):3351-3363
Polyhydroxyalkanoate depolymerase (PHAD) can be used for the degradation and recovery of polyhydroxyalkanoate (PHA). In order to develop a PHAD with good stability under high temperature, PHAD from Thermomonospora umbrina (TumPHAD) was heterelogously expressed in Escherichia coli BL21(DE3). At the same time, a mutant A190C/V240C with enhanced stability was obtained via rational design of disulfide bonds. Characterization of enzymatic properties showed that the mutant A190C/V240C had an optimum temperature of 60 ℃, which was 20 ℃ higher than that of the wild type. The half-life at 50 ℃ was 7 hours, at 50 ℃ which was 21 times longer than that of the wild type. The mutant A190C/V240C was used for the degradation of polyhydroxybutyrate (PHB), one of the typical PHA. At 50 ℃, the degradation rate of PHB being treated for 2 hours and 12 hours was 2.1 times and 3.8 times higher than that of the wild type, respectively. The TumPHAD mutant A190C/V240C obtained in this study shows tolerance to high temperature resistance, good thermal stability and strong PHB degradation ability, which may facilitate the degradation and recovery of PHB.
Thermomonospora
;
Actinomycetales
;
Escherichia coli/genetics*
;
Polyhydroxyalkanoates
3.Biodegradation of polyethylene terephthalate: a review.
Yufeng JIN ; Jiarong QIU ; Liangqing ZHANG ; Menglei ZHU
Chinese Journal of Biotechnology 2023;39(11):4445-4462
Plastics are widely used in human daily life, which bring great convenience. Nevertheless, the disposal of a large amount of plastic wastes also brings great pressure to the environment. Polyethylene terephthalate (PET) is a polymer thermoplastic material produced from petroleum. It has become one of the most commonly used plastics in the world due to its durability, high transparency, light weight and other characteristics. PET can exist in nature for a long time due to its complex structure and the difficulty in degradation, which causes serious pollution to the global ecological environment, and threatens human health. The degradation of PET wastes has since become one of the global challenges. Compared with physical and chemical methods, biodegradation is the greenest way for treating PET wastes. This review summarizes the recent advances on PET biodegradation including microbial and enzymatic degradation of PET, biodegradation pathway, biodegradation mechanisms, and molecular modification of PET-degrading enzymes. In addition, the prospect for achieveing efficient degradation of PET, searching and improving microorganisms or enzymes that can degrade PET of high crystallinity are presented, with the aimto facilitate the development, application and molecular modification of PET biodegradation microorganisms or enzymes.
Humans
;
Polyethylene Terephthalates/metabolism*
;
Polymers
;
Biodegradation, Environmental
;
Petroleum
4.An examination of the carbon metabolic pathways in Acinetobacter sp. TAC-1 in the context of poly (3-hydroxybutyrate-co-3-hydroxyvalerate) utilization.
Huan LIU ; Wang CHEN ; Senwen TAN ; Siyu LIANG ; Chenxi YANG ; Qian ZHANG
Chinese Journal of Biotechnology 2023;39(11):4663-4681
The present study aimed to unravel the carbon metabolism pathway of Acinetobacter sp. TAC-1, a heterotrophic nitrification-aerobic denitrification (HN-AD) strain that utilizes poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) as a carbon source. Sodium acetate was employed as a control to assess the gene expression of carbon metabolic pathways in the TAC-1 strain. The results of genome sequencing demonstrated that the TAC-1 strain possessed various genes encoding carbon metabolic enzymes, such as gltA, icd, sucAB, acs, and pckA. KEGG pathway database analysis further verified the presence of carbon metabolism pathways, including the glycolytic pathway (EMP), pentose phosphate pathway (PPP), glyoxylate cycle (GAC), and tricarboxylic acid (TCA) cycle in the TAC-1 strain. The differential expression of metabolites derived from distinct carbon sources provided further evidence that the carbon metabolism pathway of TAC-1 utilizing PHBV follows the sequential process of PHBV (via the PPP pathway)→gluconate (via the EMP pathway)→acetyl-CoA (entering the TCA cycle)→CO2+H2O (generating electron donors and releasing energy). This study is expected to furnish a theoretical foundation for the advancement and implementation of novel denitrification processes based on HN-AD and solid carbon sources.
3-Hydroxybutyric Acid
;
Carbon/metabolism*
;
Polyesters
;
Hydroxybutyrates
;
Metabolic Networks and Pathways
5.Research progress on medical devices of polyhydroxyalkanoate in orthopedics.
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(8):909-917
OBJECTIVE:
To review the research progress of natural biomaterial polyhydroxyalkanoate (PHA) in orthopedics.
METHODS:
The literature concerning PHA devices for bone defects, bone repair, and bone neoplasms, respectively, in recent years was extensively consulted. The three aspects of the advantages of PHA in bone repair, the preparation of PHA medical devices for bone repair and their application in orthopedics were discussed.
RESULTS:
Due to excellent biodegradability, biocompatibility, and potential osteoinduction, PHA is a kind of good bone repair material. In addition to the traditional PHA medical implants, the use of electrostatic spinning and three-dimensional printing can be designed to various functional PHA medical devices, in order to meet the orthopedic clinical demands, including the bone regeneration, minimally invasive bone tissue repair by injection, antibacterial bone repair, auxiliary establishment of three-dimensional bone tumor model, directed osteogenic differentiation of stem cells, etc.
CONCLUSION
At present, PHA is a hotspot of biomaterials for translational medicine in orthopedics. Although they have not completely applied in the clinic, the advantages of repair in bone defects have been gradually reflected in tissue engineering, showing an application prospect in orthopedics.
Orthopedics
;
Osteogenesis
;
Arthrodesis
;
Anti-Bacterial Agents
;
Biocompatible Materials
;
Polyhydroxyalkanoates/therapeutic use*
6.Preparation of functional polyhydroxyalkanoate microspheres and their antibacterial activity and osteogenic effect evaluation.
Jianfei WU ; Binglong WANG ; Yu LIU ; Daixu WEI
Chinese Journal of Reparative and Reconstructive Surgery 2023;37(8):929-936
OBJECTIVE:
To construct polyhydroxyalkanoate (PHA) microspheres loaded with bone morphogenetic protein 2 (BMP-2) and human β-defensin 3 (HBD3), and evaluate the antibacterial activity of microspheres and the effect of promoting osteogenic differentiation, aiming to provide a new option of material for bone tissue engineering.
METHODS:
The soybean lecithin (SL)-BMP-2 and SL-HBD3 were prepared by SL-mediated introduction of growth factors into polyesters technology, and the functional microsphere (f-PMS) containing BMP-2 and HBD3 were prepared by microfluidic technology, while pure microsphere (p-PMS) was prepared by the same method as the control. The morphology of microspheres was observed by scanning electron microscopy and the water absorption was detected; the release curves of BMP-2 and HBD3 in f-PMS were detected by ELISA kit. The antibacterial effect of microspheres in Staphylococcus aureus and Escherichia coli was tested with the LIVE/DEADTM BacLightTM bacterial staining kit; the biocompatibility of microspheres was tested using Transwell and cell counting kit 8 (CCK-8). The effect of microspheres on osteogenic differentiation was determined by collagen type Ⅰ (COL-1) immunofluorescence staining and alkaline phosphatase (ALP) concentration.
RESULTS:
In this experiment, the f-PMS and p-PMS were successfully constructed. Morphological characteristics showed that p-PMS surface was rough and distributed with micropores of 1-3 μm, while f-PMS surface was smooth and existed white granular material. There was no significant difference in water absorption between the two groups (P>0.05). The release curves of BMP-2 and HBD3 in the f-PMS and p-PMS were basically the same, showing both early sudden release and late slow release. The antibacterial activity of f-PMS was significantly higher than that of p-PMS in the test that against Staphylococcus aureus and Escherichia coli (P<0.05), but there was no significant difference in biocompatibility between the two groups (P>0.05). The results of osteogenic differentiation of human BMSCs showed that the fluorescence intensity of osteogenic specific protein COL-1 of f-PMS was significantly higher than that in p-PMS, and the activity of ALP in f-PMS was also significantly higher than that in p-PMS (P<0.05).
CONCLUSION
The p-PHA have good antibacterial activity and biocompatibility, and can effectively promote the osteogenic differentiation of human BMSCs, which is expected to be applied to bone tissue engineering in the future.
Humans
;
Osteogenesis
;
Polyhydroxyalkanoates
;
Microspheres
;
Alkaline Phosphatase
;
Anti-Bacterial Agents/pharmacology*
;
Coloring Agents
;
Escherichia coli
7.In Vitro Degradation Behavior of Absorbable Interface Screws.
Xuezhen ZHU ; Weizhi LIU ; Zhenlong SUN ; Shunjie YAN ; Hua LIU ; Zhongli WANG
Chinese Journal of Medical Instrumentation 2023;47(6):598-601
The composite material PLGA compounded with β-tricalcium phosphate (β-TCP) was prepared by melt blending method, and the absorbable interface screw was prepared by injection molding process. Prepare PBS buffer that simulates human body, conduct in vitro degradation experiments on interface screws according to relevant national and industry standards, then test and characterize interface screws at different time points for degradation of intrinsic viscosity, average molecular weight distribution, mass loss, mechanical properties and thermal properties. According to the degradation performance-time curve, determine the time node at which the interface screw loses the mechanical properties. In this paper, the in vitro degradation behavior of interfacial screws prepared from PLGA and β-TCP composites was studied in detail, providing a reference and basis for the degradation behavior of absorbable products prepared from PLGA and β-TCP composites.
Humans
;
Polyesters
;
Materials Testing
;
Calcium Phosphates
;
Absorbable Implants
8.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
9.Synthesis, biodegradation and waste disposal of polylactic acid plastics: a review.
Bin XIE ; Rongrong BAI ; Huashan SUN ; Xiaoli ZHOU ; Weiliang DONG ; Jie ZHOU ; Min JIANG
Chinese Journal of Biotechnology 2023;39(5):1912-1929
With the escalation of plastic bans and restrictions, bio-based plastics, represented by polylactic acid (PLA), have become a major alternative to traditional plastics in the current market and are unanimously regarded as having potential for development. However, there are still several misconceptions about bio-based plastics, whose complete degradation requires specific composting conditions. Bio-based plastics might be slow to degrade when it is released into the natural environment. They might also be harmful to humans, biodiversity and ecosystem function as traditional petroleum-based plastics do. In recent years, with the increasing production capacity and market size of PLA plastics in China, there is an urgent need to investigate and further strengthen the management of the life cycle of PLA and other bio-based plastics. In particular, the in-situ biodegradability and recycling of hard-to-recycle bio-based plastics in the ecological environment should be focused. This review introduces the characteristics, synthesis and commercialization of PLA plastics, summarizes the current research progress of microbial and enzymatic degradation of PLA plastics, and discusses their biodegradation mechanisms. Moreover, two bio-disposal methods against PLA plastic waste, including microbial in-situ treatment and enzymatic closed-loop recycling, are proposed. At last, the prospects and trends for the development of PLA plastics are presented.
Humans
;
Ecosystem
;
Biodegradable Plastics
;
Polyesters
;
Biodegradation, Environmental
10.Isolation and identification of a polyester-polyurethane degrading bacterium Bacillus altitudinis YX8-1.
Caiting ZENG ; Junbin JI ; Fanghui DING ; Zhoukun LI ; Hui CAO ; Zhongli CUI ; Xin YAN
Chinese Journal of Biotechnology 2023;39(5):1976-1986
Although polyurethane (PUR) plastics play important roles in daily life, its wastes bring serious environmental pollutions. Biological (enzymatic) degradation is considered as an environmentally friendly and low-cost method for PUR waste recycling, in which the efficient PUR-degrading strains or enzymes are crucial. In this work, a polyester PUR-degrading strain YX8-1 was isolated from the surface of PUR waste collected from a landfill. Based on colony morphology and micromorphology observation, phylogenetic analysis of 16S rDNA and gyrA gene, as well as genome sequence comparison, strain YX8-1 was identified as Bacillus altitudinis. The results of high performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) showed that strain YX8-1 was able to depolymerize self-synthesized polyester PUR oligomer (PBA-PU) to produce a monomeric compound 4, 4'-methylene diphenylamine. Furthermore, strain YX8-1 was able to degrade 32% of the commercialized polyester PUR sponges within 30 days. This study thus provides a strain capable of biodegradation of PUR waste, which may facilitate the mining of related degrading enzymes.
Polyurethanes/chemistry*
;
Polyesters/chemistry*
;
Chromatography, Liquid
;
Phylogeny
;
Tandem Mass Spectrometry
;
Bacteria/metabolism*
;
Biodegradation, Environmental

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