Ultrastructural changes of Haematococcus pluvialis (Chlorophyta) in process of astaxanthin accumulation and cell damage under condition of high light with acetate

He, Bangxiang,Hou, Lulu,Zhang, Feng,Cong, Xiaomei,Wang, Zhendong,Guo, Yalin,Shi, Jiawei,Jiang, Ming,Zhang, Xuecheng,Zang, Xiaonan The Korean Society of Phycology 2020 ALGAE Vol.35 No.3

Haematococcus pluvialis is a commercial microalga that can produce high quantities of astaxanthin. Under induced conditions, some important changes in the subcellular structures related to astaxanthin accumulation were observable. For example, a large number of astaxanthin granules, oil structures and starch granules appeared in the thick-walled cells; Astaxanthin granules gradually dissolved into the oil structures and spread throughout the entire cell with the fusion and diffusion process of oil structures during the middle and late stages of induction; The plastoglobules were closed to the newly formed structures, and some plastoglobules would abnormally increase in size under stress. Based on observations of cell damage, the degradation of membrane structures, such as chloroplasts, was found to be the primary form of damage during the early stage of induction. During the middle stage of induction, some transparent holes were exposed in the dissolving astaxanthin granules in the cytoplasm. In thick-walled cells, these transparent holes were covered by oil substances dissolving astaxanthin, thereby avoiding further damage to cells. Given the relatively few oil structures, in non-thick-walled cells, the transparent holes expanded to form multiple transparent areas, eventually resulting in the rupture and death of cells. These results suggested that the high level of synthesis and the wide range diffusion of oil explained the expansion of astaxanthin in H. pluvialis.

Pyridoxal phosphate homeostasis protein FnYggS from Fusobacterium nucleatum: purification, crystallization, and X-ray crystallographic analysis

Shanru He,Chenyuan Yuan,Xue Bai,Tingting Bu,Jie Zhang,Lulu Wang,Chunshan Quan,Yongbin Xu 한국구조생물학회 2022 Biodesign Vol.10 No.2

Anaerobic gram-negative bacterium Fusobacterium nucleatum (F. nucleatum) has long been found to cause opportunistic infections and has recently been implicated in colorectal cancer. In F. nucleatum, YggS (FnYggS) is an important part of bacterial cell walls and belongs to the COG0325 gene family. In this study, YggS from FnYggS was successfully expressed and purified using Ni-NTA affinity and gel-filtration chromatography. The protein crystal was obtained and diffracted to a resolution of 2.08 Å. The preliminary crystallographic analysis suggested that FnYggS crystal belongs to the monoclinic space group P21 with a = 37.93 Å, b = 146.38 Å, and c = 74.13 Å, α = γ = 90.00° and β = 93.36°. The asymmetric unit contained approximately three monomer of FnYggS, giving a crystal volume per mass (VM) of 2.64 Å3 Da–1 and a solvent content of 53.50%.

Overexpressing fusion proteins of 4-coumaroyl-CoA ligase (4CL) and stilbene synthase (STS) in tobacco plants leading to resveratrol accumulation and improved stress tolerance

Xuancheng He,Feiyan Xue,Lulu Zhang,Hui-Li Guo,Lan-Qing Ma,Mingfeng Yang 한국식물생명공학회 2018 Plant biotechnology reports Vol.12 No.5

Resveratrol (trans-3,4′,5-trihydroxystilbene) is a phytoalexin produced in plants in response to pathogen attack as a part of plant defense response and it is also a highly bioactive substance of pharmaceutical interest. To obtain transgenic plants with a high level of resveratrol, two enzymes in the last two steps of resveratrol synthesis, 4-coumaroyl-CoA ligase (4CL) and stilbene synthase (STS), were fused together by a glycine–serine–glycine (GSG) tripeptide linker, and the 4CL-GSGSTS construct driven by a CaMV35S promoter was transformed into tobacco (Nicotiana benthamiana) by Agrobacteriummediated method. In the transgenic plants, a high resveratrol level was detected (21.05 μg/g fresh weight) by high-pressure liquid chromatography (HPLC), which is higher than previous transgenic plants with only STS gene overexpression. In addition to resistance to pathogen, transgenic plants showed improved tolerance to salt and osmotic stresses, and the lower level of malondialdehyde (MDA) in transgenic plants suggested that resveratrol could protect plant membrane lipid from peroxidation under abiotic stresses.

Environmentally Friendly Waterproof and Breathable Nanofiber Membranes with Thermal Regulation Performance by One-Step Electrospinning

Pan Zhang,Gaihuan Ren,Lulu Tian,Bo Li,Zhanzhao Li,Hongqin Yu,Rongwu Wang,Jianxin He 한국섬유공학회 2022 Fibers and polymers Vol.23 No.8

The waterproof and moisture permeable fabric with thermal regulation not only has the protective function forhuman body, but also meets the comfort of human clothing, which is a challenging and innovative research. Herein, we reportan environmentally friendly, simple, and high-efficiency strategy to fabricate waterproof and breathable nanofibermembranes with outstanding thermal regulation ability by introducing stearic acid (SA) into the hydrophobic siliceouspolyurethane (SIPU) solution by a facile one-step electrospinning. Owing to the addition of SA, the waterproofness of SIPU/SA nanofiber membranes is greatly improved, meanwhile, the addition of SA acted as a phase change material to imparttemperature regulation performance in appropriate temperature range to the SIPU/SA nanofiber membranes. By adjusting theaddition amount of the SA, eco-friendly and multifunctional SIPU/SA nanofiber membranes are systemically optimized withgood waterproofness (79 kPa), moisture breathability (8.0 kg m-2 d-1), and outstanding temperature regulation performance(latent heat up to 40 J/g), satisfied tensile strength (6.2 MPa) and excellent breaking elongation of 182 %. The obtained SIPU/SA nanofiber membranes can be suitable for the design of multifunctional wearable textiles for various applications,especially in outdoor protective clothing, medical clothing, intelligent clothing and military products.

Fibroblast Growth Factor 21 Attenuates Diabetes- Induced Renal Fibrosis by Negatively Regulating TGF- β-p53-Smad2/3-Mediated Epithelial-to-Mesenchymal Transition via Activation of AKT

Sundong Lin,Lechu Yu,Yongqing Ni,Lulu He,Xiaolu Weng,Xuemian Lu,Chi Zhang 대한당뇨병학회 2020 Diabetes and Metabolism Journal Vol.44 No.1

Background: Epithelial-to-mesenchymal transition (EMT) is required for renal fibrosis, which is a characteristic of diabetic nephropathy (DN). Our previous study demonstrated that fibroblast growth factor 21 (FGF21) prevented DN associated with the suppressing renal connective tissue growth factor expression, a key marker of renal fibrosis. Therefore, the effects of FGF21 on renal fibrosis in a DN mouse model and the underlying mechanisms were investigated in this study. Methods: Type 1 diabetes mellitus was induced in C57BL/6J mice by intraperitoneal injections of multiple low doses of streptozotocin. Then, diabetic and non-diabetic mice were treated with or without FGF21 in the presence of pifithrin-α (p53 inhibitor) or 10-[4´-(N,N-Diethylamino)butyl]-2-chlorophenoxazine hydrochloride (10-DEBC) hydrochloride (Akt inhibitor) for 4 months. Results: DN was diagnosed by renal dysfunction, hypertrophy, tubulointerstitial lesions, and glomerulosclerosis associated with severe fibrosis, all of which were prevented by FGF21. FGF21 also suppressed the diabetes-induced renal EMT in DN mice by negatively regulating transforming growth factor beta (TGF-β)-induced nuclear translocation of Smad2/3, which is required for the transcription of multiple fibrotic genes. The mechanistic studies showed that FGF21 attenuated nuclear translocation of Smad2/3 by inhibiting renal activity of its conjugated protein p53, which carries Smad2/3 into the nucleus. Moreover pifithrin-α inhibited the FGF21-induced preventive effects on the renal EMT and subsequent renal fibrosis in DN mice. In addition, 10-DEBC also blocked FGF21-induced inhibition of renal p53 activity by phosphorylation of mouse double minute-2 homolog (MDM2). Conclusion: FGF21 prevents renal fibrosis via negative regulation of the TGF-β/Smad2/3-mediated EMT process by activation of the Akt/MDM2/p53 signaling pathway.

Effect of Rapid Chilling on Beef Quality and Cytoskeletal Protein Degradation in M. longissimus of Chinese Yellow Crossbred Bulls

Mao, Yanwei,Zhang, Yimin,Liang, Rongrong,Ren, Lulu,Zhu, He,Li, Ke,Zhu, Lixian,Luo, Xin Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.8

The objective of this study was to investigate the effect of rapid chilling (RC) on beef quality and the degradation of cytoskeletal proteins. Twenty Chinese Yellow crossbred bulls were selected and randomly divided into two groups. RC and conventional chilling (CC) were applied to left and right sides of the carcasses respectively after slaughtering. To determine whether electrical stimulation (ES) treatment can alleviate the potential hazard of RC on meat quality, ES was applied to one group. The effects of RC and ES were determined by meat color, shear force and cytoskeletal protein degradation postmortem (PM). The results showed that RC decreased beef tenderness at 1 d and 3 d postmortem, but had no detrimental effect on meat color. Western blotting showed that RC decreased the degradation rate of desmin and troponin-T, but the effects weakened gradually as postmortem aging extended. Degradation rates of both desmin and troponin-T were accelerated by ES. The combination of RC and ES could improve beef color, accelerate degradation rate of cytoskeletal protein and improve beef tenderness.

Therapeutic Effects of Fibroblast Growth Factor-21 on Diabetic Nephropathy and the Possible Mechanism in Type 1 Diabetes Mellitus Mice

Weng Wenya,Ge Tingwen,Wang Yi,He Lulu,Liu Tinghao,Wang Wanning,Zheng Zongyu,Yu Lechu,Zhang Chi,Lu Xuemian 대한당뇨병학회 2020 Diabetes and Metabolism Journal Vol.44 No.4

Background: Fibroblast growth factor 21 (FGF21) has been only reported to prevent type 1 diabetic nephropathy (DN) in the streptozotocin-induced type 1 diabetes mellitus (T1DM) mouse model. However, the FVB (Cg)-Tg (Cryaa-Tag, Ins2-CALM1) 26OVE/PneJ (OVE26) transgenic mouse is a widely recommended mouse model to recapture the most important features of T1DM nephropathy that often occurs in diabetic patients. In addition, most previous studies focused on exploring the preventive effect of FGF21 on the development of DN. However, in clinic, development of therapeutic strategy has much more realistic value compared with preventive strategy since the onset time of DN is difficult to be accurately predicted. Therefore, in the present study OVE26 mice were used to investigate the potential therapeutic effects of FGF21 on DN. Methods: Four-month-old female OVE26 mice were intraperitoneally treated with recombinant FGF21 at a dose of 100 μg/kg/day for 3 months. The diabetic and non-diabetic control mice were treated with phosphate-buffered saline at the same volume. Renal functions, pathological changes, inflammation, apoptosis, oxidative stress and fibrosis were examined in mice of all groups. Results: The results showed that severe renal dysfunction, morphological changes, inflammation, apoptosis, and fibrosis were observed in OVE26 mice. However, all the renal abnormalities above in OVE26 mice were significantly attenuated by 3-month FGF21 treatment associated with improvement of renal adenosine 5´-monophosphate (AMP)-activated protein kinase (AMPK) activity and sirtuin 1 (SIRT1) expression. Conclusion: Therefore, this study demonstrated that FGF21 might exert therapeutic effects on DN through AMPK-SIRT1 pathway.

NADH-dependent butanol dehydrogenase from Fusobacterium nucleatum: purification, crystallization, and X-ray crystallographic analysis

Xue Bai,Jing Lan,Shanru He,Tingting Bu,Jie Zhang,Lulu Wang,Chunshan Quan,Ki Hyun Nam,Nam-Chul Ha,Yongbin Xu 한국구조생물학회 2022 Biodesign Vol.10 No.2

Fusobacterium nucleatum is a dangerous pathogen, and it has been linked to a variety of health problems, including cancer, periodontal disease, and pregnancy complications. F. nucleatum butanol dehydrogenase (FnYqdH) is a group of dehydrogenase enzymes that facilitate interconversion between butyraldehyde and butanol at the expenditure of a cofactor NAD (P)H. In this study, FnYqdH was successfully expressed and purified using Ni-NTA affinity, Q anionexchange, and gel-filtration chromatography. The protein crystal was obtained and diffracted to a resolution of 2.0 Å. The crystal belonged to the orthorhombic space group of I222, with unit-cell parameters of a = 64.77, b = 78.85, and c = 215.22 Å. The Matthews coefficient and solvent content were estimated to be 3.19 Å3 Da–1 and 61.49%, respectively, assuming that the asymmetric unit contained only one recombinant protein molecule. Size-exclusion chromatography suggested that FnYqdH prefers to exist as a dimer in the solution.

Clostridium beijerinckii glyceraldehyde-3-phosphate dehydrogenase GAPDH: purification, crystallization, and X-ray crystallographic analysis

Jie Zhang,Yuanyuan Chen,Tingting Bu,Xue Bai,Shanru He,Lulu Wang,Chunshan Quan,Yongbin Xu 한국구조생물학회 2022 Biodesign Vol.10 No.3

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a ubiquitous and highly abundant glycolytic enzyme. It plays a pivotal role for the energy and carbon metabolism of most organisms including industrial bacteria. It catalyzes the two step oxidative phosphorylation of D-glyceraldehyde-3-phosphate into 1,3-bisphosphoglycerate using inorganic phosphate and nicotinamide adenine dinucleotide (NAD+) as cofactor. In this study, GAPDH from C. beijerinckii (CbGAPDH) was successfully expressed and purified using Ni-NTA affinity, Q anion-exchange, and gel-filtration chromatography. The protein crystal was obtained and diffracted to a resolution of 1.60 Å. The crystal belonged to the hexagonal space group P6222, with unit-cell parameters of a = 120.6, b = 120.6, and c = 122.1 Å. The Matthews coefficient and solvent content were estimated to be 3.50 Å3 Da–1 and 64.90%, respectively, assuming that the asymmetric unit contained only one recombinant protein molecule.