Journal Articles
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Item Impact of cell wall polysaccharide modifications on the performance of Pichia pastoris: novel mutants with enhanced fitness and functionality for bioproduction applications.(BioMed Central Ltd, 2024-02-17) Cheng B; Yu K; Weng X; Liu Z; Huang X; Jiang Y; Zhang S; Wu S; Wang X; Hu XBACKGROUND: Pichia pastoris is a widely utilized host for heterologous protein expression and biotransformation. Despite the numerous strategies developed to optimize the chassis host GS115, the potential impact of changes in cell wall polysaccharides on the fitness and performance of P. pastoris remains largely unexplored. This study aims to investigate how alterations in cell wall polysaccharides affect the fitness and function of P. pastoris, contributing to a better understanding of its overall capabilities. RESULTS: Two novel mutants of GS115 chassis, H001 and H002, were established by inactivating the PAS_chr1-3_0225 and PAS_chr1-3_0661 genes involved in β-glucan biosynthesis. In comparison to GS115, both modified hosts exhibited a looser cell surface and larger cell size, accompanied by faster growth rates and higher carbon-to-biomass conversion ratios. When utilizing glucose, glycerol, and methanol as exclusive carbon sources, the carbon-to-biomass conversion rates of H001 surpassed GS115 by 10.00%, 9.23%, and 33.33%, respectively. Similarly, H002 exhibited even higher increases of 32.50%, 12.31%, and 53.33% in carbon-to-biomass conversion compared to GS115 under the same carbon sources. Both chassis displayed elevated expression levels of green fluorescent protein (GFP) and human epidermal growth factor (hegf). Compared to GS115/pGAPZ A-gfp, H002/pGAPZ A-gfp showed a 57.64% higher GFP expression, while H002/pPICZα A-hegf produced 66.76% more hegf. Additionally, both mutant hosts exhibited enhanced biosynthesis efficiencies of S-adenosyl-L-methionine and ergothioneine. H001/pGAPZ A-sam2 synthesized 21.28% more SAM at 1.14 g/L compared to GS115/pGAPZ A-sam2, and H001/pGAPZ A-egt1E obtained 45.41% more ERG at 75.85 mg/L. The improved performance of H001 and H002 was likely attributed to increased supplies of NADPH and ATP. Specifically, H001 and H002 exhibited 5.00-fold and 1.55-fold higher ATP levels under glycerol, and 6.64- and 1.47-times higher ATP levels under methanol, respectively, compared to GS115. Comparative lipidomic analysis also indicated that the mutations generated richer unsaturated lipids on cell wall, leading to resilience to oxidative damage. CONCLUSIONS: Two novel P. pastoris chassis hosts with impaired β-1,3-D-glucan biosynthesis were developed, showcasing enhanced performances in terms of growth rate, protein expression, and catalytic capabilities. These hosts exhibit the potential to serve as attractive alternatives to P. pastoris GS115 for various bioproduction applications.Item Evolutionary Game and Simulation of Green Housing Market Subject Behavior in China.(John Wiley and Sons, 2022-04-05) Qian Y; Yu M; Wang T; Yuan R; Feng Z; Zhao X; Fu HIn China, driven by the national "3060" double carbon targets (i.e., reaching peak carbon emissions by 2030 and carbon neutrality by 2060), green housing has become one of the major fields to reduce carbon emissions, facilitating the achievement of the double carbon targets. Promoting the growth of green housing is an important way for the real estate industry to achieve low-carbon transformation and improve the quality of housing. Meanwhile, the construction industry also can benefit from green housing to achieve its energy conservation and emission reduction targets. Therefore, it is critical to boost and maintain the sustainable growth of the green housing market in China. However, the literature has not focused attention on the market behavior of the green housing market in China. This study proposes a tripartite evolutionary game model to investigate the subject behavior of the green housing market in China. This model consists of three major subjects in a green housing market: developers, consumers, and governments. Based on this model, this study analyzes the stability of the strategy options for each stakeholder and identifies the stable conditions of strategy portfolios to reach the equilibrium points of the game system. The validity of the proposed tripartite evolutionary game model is tested through the simulation of the impacts from various factors on system evolution. According to the impacts of factors and the stable conditions of strategies, this paper puts forward relevant policy suggestions for the healthy and sustainable growth of China's green housing market.Item Role of a local transcription factor in governing cellular carbon/nitrogen homeostasis in Pseudomonas fluorescens(Oxford University Press on behalf of Nucleic Acids Research, 2021-04-06) Naren N; Zhang X-XAutoactivation of two-component systems (TCSs) can increase the sensitivity to signals but inherently cause a delayed response. Here, we describe a unique negative feedback mechanism enabling the global NtrB/NtrC regulator to rapidly respond to nitrogen starvation over the course of histidine utilization (hut) in Pseudomonas fluorescens. NtrBC directly activates transcription of hut genes, but overexpression will produce excess ammonium leading to NtrBC inactivation. To prevent this from occurring, the histidine-responsive repressor HutC fine-tunes ntrBC autoactivation: HutC and NtrC bind to the same operator site in the ntrBC promoter. This newly discovered low-affinity binding site shows little sequence similarity with the consensus sequence that HutC recognizes for substrate-specific induction of hut operons. A combination of genetic and transcriptomic analysis indicated that both ntrBC and hut promoter activities cannot be stably maintained in the ΔhutC background when histidine fluctuates at high concentrations. Moreover, the global carbon regulator CbrA/CbrB is involved in directly activating hut transcription while de-repressing hut translation via the CbrAB-CrcYZ-Crc/Hfq regulatory cascade. Together, our data reveal that the local transcription factor HutC plays a crucial role in governing NtrBC to maintain carbon/nitrogen homeostasis through the complex interactions between two TCSs (NtrBC and CbrAB) at the hut promoter.Item Hagfish feeding habits along a depth gradient inferred from stable isotopes(27/06/2013) Zintzen V; Rogers KM; Roberts CD; Stewart AL; Anderson MJFeeding habits of 3 hagfish species were investigated along a depth gradient (~50 to 900 m) in New Zealand using nitrogen (?15N) and carbon (?13C) stable isotopes. Neomyxine biniplicata had the lowest mean ?15N value (14.2%), followed by Eptatretus cirrhatus (14.9 %) and Eptatretus sp. 1 (15.8%). Neomyxine biniplicata (~50 m depth) was characterized by (1) relative low lipid content in muscles and (2) consistent body condition index which together with its trophic position indicated that this species probably acquires its food by active predation, supplemented by opportunistic scavenging. Eptatretus cirrhatus (48 to 912 m) and Eptatretus sp. 1 (290 to 922 m) had similar morphology, but their ?15N signature indicated that they were feeding on slightly different trophic levels. For Eptatretus sp. 1, the combination of (1) variable lipid content, indicating phases of feeding and fasting, (2) decreasing body condition index with depth, indicating less regular feeding at depth, (3) increasing ?15N with depth and (4) decreasing ?13C signature with depth, pointed towards a feeding behaviour specialized in scavenging on large but rare falls of high-level predators such as whales, sharks or bony fishes. On the other hand, E. cirrhatus was characterized by (1) less variable lipid content, (2) a body condition index not influenced by depth, (3) ?15N values decreasing with depth and ?13C values constant across its depth range, which is likely to indicate a more opportunistic and mobile feeding behaviour on a range of prey © 2013 Museum of New Zealand Te Papa Tongarewa and Massey University and GNS Science.
