关键词： Halomonas   Synthetic biology   PHB   4-Hydroxybutyrate   3-Hydroxyvalerate   Next generation industrial biotechnology   NGIB  

摘要： Microbial poly(3-hydroxybutyrate-co-4-hydroxybutyrate-co-3-hydroxyvalerate), abbreviated as P(3HB-4HB3HV) or P34HBHV, is a flexible polyhydroxyalkanoate (PHA) material ranging from softness to elasticity depending on the ratios of various monomers. Halomonas bluephagenesis, as the chassis of the next generation industrial biotechnology (NGIB) able to grow contamination free under open unsterile conditions. The resulting recombinants of H. bluephagenesis became capable of efficiently synthesizing P34HBHV utilizing glucose as the sole carbon source. Engineered H. bluephagenesis H1 (encoding ogdA, sucD, 4hbD, orfZ, scpA and scpB in chromosomes) transformed with a plasmid containing PHA synthesis genes phaC and phaA and its derivative H29 produced up to 92 % P(3HB-co-8.85 %4HB-co-8.47 %3HV) and 72 % P(3HB-co-13.21 %4HB-co-11.97 %3HV) in cell dry weight (CDW), respectively, in shake flasks. In bioreactor cultivation, H. bluephagenesis H39 constructed by integrating the 4hbD, phaC and phaA genes into the genome of H. bluephagenesis H1 achieved 95 g/L CDW with 69 % P(3HB-co-10.49 %4HB-co-3.54 %3HV), while H. bluephagenesis H43, further optimized with lpxM deletion, reached 73 g/L CDW with 78 % P(3HB-co-10.35 %4HB-co-4.54 %3HV) in a 100 L bioreactor. For the first time, H. bluephagenesis was successfully engineered to generate stable and hyperproductive derivative strains for pilot production of P(3HB-4HB-3HV) with customizable monomer ratios from glucose as the sole carbon source.

关键词： Cartilage decellularized ECM   Polyhydroxybutyrate   Chitosan   Halloysite nanotubes   Core-shell   Electrospun scaffold  

摘要： Cartilage regeneration is still a highly challenging field due to its low self-healing ability. This study used a coreshell electrospinning technique to enhance cartilage tissue engineering by incorporating cartilage extracellular matrix (ECM). The core of fibers included poly(3-hydroxybutyrate)-Chitosan (PHB-Cs) and Halloysite nanotubes. The shell of fibers consisted of Cs and ECM (0, 1, 3, 5 wt%). Subsequently, the scaffolds were named 0E, 1E, 3E, and 5E. The study aimed to assess the impact of ECM on cellular behavior and chondrogenesis. Our findings indicate that ECM reduced fiber diameter from 775 nm for the 0E scaffold to 454 nm for the 1E scaffold. Water contact angle measurements revealed an increasing trend by ECM addition, from 42 degrees for 0E to 67 degrees for 1E. According to mechanical analysis, the 1E scaffold represented the highest strength (5.81 MPa) and strain (3.17%). Based on these analyses, the 1E was considered the optimum scaffold. MTT analysis showed cell viability of over 80% for the 0E and 1E. Also, the gene expression level was assessed for Collagen II, Aggrecan, SOX 9, and Collagen X. The results represented that in the 1E scaffold Collagen II, Aggrecan, and SOX 9 were more upregulated at the end of the 21st day. However, in the 1E scaffold collagen X, as a hypertrophy marker, was downregulated at the end of the experiment. Overall, these results confirmed the potential of the 1E scaffold to be introduced as a promising cartilage tissue engineering scaffold for further studies.

关键词： Bioplastics   Lignocellulose   Next generation industrial biotechnology   Xylose   PHB   PHA   Metabolic engineering  

摘要： Lignocellulose is the most abundant terrestrial biomass type, and lignocellulose hydrolysate has the potential to replace glucose for microbial fermentation. Halomonas bluephagenesis has significant advantages in producing bioplastics polyhydroxyalkanoates (PHA), but there is relatively little research on the use of lignocellulose hydrolysate for this strain. In present study, H. bluephagenesis was engineered to use xylose and lignocellulose hydrolysate to produce PHB. Firstly, four xylose metabolism pathways were established. Secondly, several xfp genes were compared and genes in pathway I (xylA and xfp gene) were integrated into the genome. Thirdly, H. bluephagenesis was found to be able to utilize glucose and xylose simultaneously. H. bluephagenesis T39 containing xylA and xfp generated 15 g/L CDW containing 76 wt% PHB when cultured in lignocellulose hydrolysate, and it was grown to 62 g/L CDW containing 67 wt% PHB in a 7 L bioreactor. H. bluephagenesis T43 harboring xylA was found able to synthesize P(3HB-4HB-3HV) containing 3-hydroxybutyrate (3HB), 4-hydroxybutyrte (4HB) and 3-hydroxyvalerate (3HV) when grown on lignocellulose hydrolysate.

关键词： PHB   Halomonas   3-Hydroxyvalerate   beta-Oxidation   NGIB   Synthetic biology  

摘要： Polyhydroxyalkanoates (PHA) with high 3-hydroxyvalerate (3HV) monomer ratios lead to their accelerated biodegradation and improved thermal and mechanical properties. In this study, poly(3-hydroxybutyrate-co-3- hydroxyvalerate) (PHBV) with a broad range of 3HV ratios were produced and characterized using the next generation industrial biotechnology (NGIB) chassis Halomonas bluephagenesis (H. bluephagenesis). Wild type H. bluephagenesis was found to produce P(3HB-co-66.31mol% 3HV) when cultured in the presence of valerate. Deletion on the functional enoyl-CoA hydratase (fadB1) increased to 93.11 mol% 3HV in the PHBV copolymers. Through tuning the glucose and valerate co-feeding, PHBV with controllable 3HV ratios were adjusted to range from 0-to-93.6 mol% in shake-flask studies. Metabolic weakening of the beta-oxidation pathway paired with flux limitation to the native 3HB synthesis pathway were used to reach the highest reported 98.3 mol% 3HV by H. bluephagenesis strain G34B grown in shake flasks. H. bluephagenesis strain G34B was grown to 71.42 g/L cell dry weight (CDW) containing 74.12 wt% P(3HB-co-17.97 mol% 3HV) in 7 L fermentors. Mechanical properties of PHBV with 0, 22.81, 42.76, 73.49 and 92.17 mol% 3HV were characterized to find not linearly related to increased 3HV ratios. Engineered H. bluephagenesis has demonstrated as a platform for producing PHBV of various properties.

关键词： 高寒地区   城镇生活污水   处理技术   生物处理   强化措施  

摘要： 高寒地区空气稀薄、寒冷干燥、紫外线辐射强，导致城镇生活污水处理效率较低且不稳定，给当地的城镇生活污水处理带来挑战，有必要对其城镇生活污水处理技术进行总结以提升处理效率。总结了我国高寒地区城镇生活污水处理技术应用现状，指出高寒地区城镇生活污水处理主要采用氧化沟、SBR、A2O、MBR系列工艺；阐释了污水处理中存在的微生物活性低、污泥膨胀、微生物丰度下降等问题；梳理出城镇生活污水处理强化措施包括调控工艺参数、改良工艺、设置保温措施、采用组合工艺、进行生物强化等。针对高寒地区低温影响生活污水处理效率问题，建议从低温下培养耐冷菌、采用SBR及MBR系列工艺、开发新工艺三方面开展研究，以强化污水处理效能；针对高寒地区低氧问题，建议从优化曝气系统、增加曝气时间、结合厌氧好氧工艺三方面开展研究。未来，绿色低碳技术的研发应用将为高寒地区城镇生活污水处理提供新的解决方案。

关键词： Neurodegeneration   parkinson disease   PHB2-LC3 interaction   PINK1-PRKN   porin   ubiquitin-proteasome system  

摘要： Exposure of inner mitochondrial membrane resident protein PHB2 (prohibitin 2) during autophagic removal of depolarized mitochondria (mitophagy) depends on the ubiquitin-proteasome system. This uncovering facilitates the PHB2 interaction with phagophore membrane-associated protein MAP1LC3/LC3. It is unclear whether PHB2 is exposed randomly at mitochondrial rupture sites. Prior knowledge and initial screening indicated that VDAC1 (voltage dependent anion channel 1) might play a role in this phenomenon. Through in vitro biochemical assays and imaging, we have found that VDAC1-PHB2 interaction increases during mitochondrial depolarization. Subsequently, this interaction enhances the efficiency of PHB2 exposure and mitophagy. To investigate the relevance in vivo, we utilized porin (equivalent to VDAC1) knockout Drosophila line. Our findings demonstrate that during mitochondrial stress, porin is essential for Phb2 exposure, Phb2-Atg8 interaction and mitophagy. This study highlights that VDAC1 predominantly synchronizes efficient PHB2 exposure through mitochondrial rupture sites during mitophagy. These findings may provide insights to understand progressive neurodegeneration.

关键词： 污水处理   装置自净   活性炭   净化效率  

摘要： 文章设计了一种简易废液处理及自净的装置,装置由净水箱、过滤箱和储水箱三大主体组成,净水箱中有活性炭吸附板起到净化污水的作用,通过喷水机构和传动机构等的配合使用,将装置自身清洗干净,避免污渍久积影响净化效率。选取郎家厂水质站汛期时水样作为研究对象,实验结果表明装置净化效果很好,净化效率均在90%左右,具有较好的发展前景。

关键词： 酒庄废水   宏基因组学   真菌   群落结构   功能特征  

摘要： 为探究酒庄污水处理系统中真菌的群落结构和功能特征,本文以银川市某酒庄SBBR污水处理系统为研究对象,利用宏基因组学测序技术,分析酒庄污水处理系统活性污泥和生物膜中真菌的菌群结构和功能特征。结果表明:SBBR污水处理系统中检出真菌10门34纲78目169科279属484种。优势真菌门为子囊菌门(Ascomycota)(34.07%~42.32%)和担子菌门(Basidiomycota)(22.82%~24.99%),优势真菌属为罗兹菌属(Rozella)(3.55%~5.69%)和微孢子虫(Mitosporidium)(3.48%~5.09%)。功能组成中与代谢相关的通路为丰度最高的通路(25.37%~38.58%),其中嘧啶代谢(Pyrimidine metabolism)(2.37%~7.44%)和嘌呤代谢(Purine metabolism)(1.77%~2.58%)为优势代谢通路,并且在活性污泥中具有比生物膜更高的功能丰度。曲霉属(Aspergillus)是构成活性污泥和生物膜组间以及功能差异的主要真菌;嘧啶代谢(Pyrimidine metabolism)是构成两者功能差异的主要代谢通路。物种与功能贡献度分析中,担子菌门(Basidiomycota)在参与双组分系统(Two-component system)功能中占据了最大贡献度,平均贡献度为80.9%。

关键词： ARES   纳米磁畴   光合细菌   组合工艺  

摘要： 漳州某污水处理厂以处理海产品加工废水为主,针对其废水水质具有成分复杂、有机污染物浓度高、氮磷高等特点,设计采用生态能污水处理系统,结合纳米磁畴震滤的深度处理工艺;文中以此工程为例,介绍了该组合工艺的流程、原理、设计参数及工艺特点等。工程运行结果表明,该组合工艺运行稳定,系统对污染物去除效率高,出水各项指标优于一级A排放标准,为同类型废水的处理提供参考。

关键词： 城镇污水处理厂   环境、社会和公司治理(ESG)   层次分析法   模糊综合评价法  

摘要： 构建了一种基于ESG+E(Environment,Social,and Governance+Economy)的城镇污水处理厂评价体系,从环境影响、社会责任、公司治理和经济效益4个维度出发,建立了7个细化准则及20个评价指标.采用层次分析法和模糊综合评价法构建评价模型,并选取上海市3座城镇污水处理厂(WWTP1、WWTP2、WWTP3)进行绩效评价.结果表明,WWTP1和WWTP3的整体绩效均处于优秀水平;WWTP2的整体绩效处于良好水平,今后可采取提高污水资源化利用率,提供多样化的员工培训机会,完善信息公开制度等改进措施以提高污水处理厂的绩效水平.3座城镇污水处理厂的评价结果与实际运行情况相符,验证了所构建评价模型的准确性和科学性,为城镇污水处理厂的绩效评价工作提供参考.