INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
·
IF 7.7
Xuanwei Mei,Wenwen Tao,Haitao Sun,Guanchen Liu,Guangning Chen,Yuying Zhang,Changhu Xue,Yaoguang Chang
Alginate is a commercially important polysaccharide widely distributed in brown algae. Carbohydrate-binding modules (CBMs), a class of commonly used polysaccharide-binding proteins, have greatly facilitated the investigations of polysaccharides. Few alginate-binding CBMs have been hitherto reported and structurally characterized. Herein, an unknown domain from a potential PL6 family alginate lyase in the marine bacterium Vibrio breoganii was discovered and recombinantly expressed. The obtained protein, designated VbCBMxx, displayed the favorable specificity to alginate. The unique sequence and well-defined function of VbCBMxx reveal a new CBM family. Moreover, the structure of VbCBMxx was determined at a 1.5 Å resolution by the X-ray crystallography, which shows a typical β-sandwich fold comprised of two antiparallel β-sheets. Site-directed mutagenesis assays confirmed that positively charged polar residues are crucial for the ligand binding of VbCBMxx. The discovery of VbCBMxx enriches the toolbox of alginate-binding proteins, and the elucidation of critical residues would guide the future practical applications of VbCBMxx.
研究领域:
生物化学
·
结构生物学
·
海洋微生物学
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DOI
PubMed
高影响因子
CARBOHYDRATE POLYMERS
·
2024
·
IF 10.7
Xueyan Wang,Le Liu,Ruiyu Shen,Qian Wang,Xiaoqi Xie,Weiming Liu,Zhimin Yu,Xianzhen Li,Xiaoyu Guo,Fan Yang
The inefficient decomposition of polysaccharides, particularly branched polysaccharides limits their large-scale industrial applications. Further understanding and modification of glycoside hydrolases (GHs) processivity is expected to overcome this limitation. Here, a novel xanthan-binding CBM ( Mi XBM), which was supposed to alter the processivity of GHs, was systematically characterized. Phylogeny and structure analyses indicated that Mi XBM is closely related to putative polysaccharide side chain-binding modules. Quantitative binding assays further revealed that Mi XBM probably has a high affinity for xanthan side chain via a variable loop site. Moreover, catalytic performance demonstrated that xanthanase chimeras containing Mi XBM promote highly efficient hydrolysis of xanthan because of improved substrate accessibility. Notably, Mi XBM was observed to enhance the processivity of xanthanase, owing to its high substrate affinity to the repeating unit xanthan. Furthermore, sequential hydrolysis of xanthan by xanthanases with varying processivity resulted in significantly increased hydrolytic efficiency and focused oligoxanthans array. These results expand understanding of CBM-substrate recognition and shed light on efficient degradation of other regularly branched polysaccharides using modified GHs.
CAS / MDL:
研究领域:
生物化学
·
分子生物学
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DOI
PubMed
高影响因子
ACS Omega
·
2020
·
IF 2.87
Xueyu Liu,Hongguang Zhu
The traditional biological denitrification process has the problems of low removal rates and lack of a carbon source when treating wastewater with high ammonia nitrogen concentration and a low carbon–nitrogen ratio. Based on a bio-electrochemical system (BES), a novel carbon cloth bipolar plate multicompartment electroenhanced bioreactor (CBM-EEB) system was constructed. In this study, nitrogen removal efficiency and enrichment of functional bacteria using CBM-EEB under different voltage conditions were investigated. The results from next-generation sequencing indicated that the CBM-EEB included heterotrophic nitrification and aerobic denitrification (HNAD) and was dominated by heterotrophic nitrification aerobic denitrifying bacteria (HNADB). The applied voltage was confirmed as having the ability to regulate the microbial community structure and abundance of functional genes, thereby further enhancing the nitrogen removal efficiency of the system. The total nitrogen removal efficiency was 77.70 ± 1.14, 87.10 ± 0.56, 86.40 ± 0.59, and 89.30 ± 0.53% under applied voltages of 0.4, 0.7, 1.0, and 1.3 V, respectively. All values were significantly higher than the control group (62.86 ± 2.06%). HNADB had the highest abundance among the 17 detected genera related to nitrogen metabolism. Facultative denitrifying bacteria, Pseudoxanthomonas, along with key bacteria of HNADB, such as Flavobacterium, constructed a shortcut simultaneous nitrification–denitrification (SND) process. Poisson analysis and redundancy analysis (RDA) showed that the applied voltage improved the denitrification efficiency by changing the microbial community structure, reducing the abundance of heterotrophic bacteria, and increasing the unit abundance of key functional genes so that less organics were required for the denitrification process. The increased nitrogen removal efficiency in the experimental group was mainly related to simultaneous nitrification–denitrification process and cooperation of microbial communities in the anode and the cathode. This study highlighted the feasibility of CBM-EEB to enhance the HNAD reaction and the response of wastewater with a low C/N ratio to enhance the abundance of microbial bacteria and their functional gene abundance.
研究领域:
环境工程
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生物技术
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微生物学
·
废水处理
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DOI
PubMed
高影响因子
BMC BIOTECHNOLOGY
·
2017
·Vol. 17
·期 1
·1-11
·
IF 2.415
Chang Fei,Zhang Xianbing,Pan Yu,Lu Youxue,Fang Wei,Fang Zemin,Xiao Yazhong
Background β-Glucosidase has attracted substantial attention in the scientific community because of its pivotal role in cellulose degradation, glycoside transformation and many other industrial processes. However, the tedious and costly expression and purification procedures have severely thwarted the industrial applications of β-glucosidase. Thus development of new strategies to express β-glucosidases with cost-effective and simple procedure to meet the increasing demands on enzymes for biocatalysis is of paramount importance.Results Light activated cassette YF1/FixJ and the SRRz lysis system were successfully constructed to produce Bgl1A(A24S/F297Y), a mutant β-glucosidase tolerant to both glucose and ethanol. By optimizing the parameters for light induction, Bgl1A(A24S/F297Y) activity reached 33.22 ± 2.0 U/mL and 249.92 ± 12.25 U/mL in 250-mL flask and 3-L fermentation tank, respectively, comparable to the controls of 34.02 ± 1.96 U/mL and 322.21 ± 10.16 U/mL under similar culture conditions with IPTG induction. To further simplify the production of our target protein, the SRRz lysis gene cassette from bacteriophage Lambda was introduced to trigger cell autolysis. As high as 84.53 ± 6.79% and 77.21 ± 4.79% of the total β-glucosidase were released into the lysate after cell autolysis in 250 mL flasks and 3-L scale fermentation with lactose as inducer of SRRz. In order to reduce the cost of protein purification, a cellulose-binding module (CBM) from Clostridium thermocellum was fused into the C-terminal of Bgl1A(A24S/F297Y) and cellulose was used as an economic material to adsorb the fusion enzyme from the lysate. The yield of the fusion protein could reach 92.20 ± 2.27% after one-hour adsorption at 25 °C.Conclusion sWe have developed an efficient and inexpensive way to produce β-glucosidase for potential industrial applications by using the combination of light induction, cell autolysis, and CBM purification strategy.
研究领域:
生物技术
·
酶工程
·
发酵工程
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DOI
PubMed
高影响因子
Cancer Biomarkers
·
2016
·Vol. 16
·期 3
·477-487
·
IF 1.736
Li Zhen,Shu Jinian,Zhang Peng,Sun Wanqi,Yang Bo,Zhang Haixu
BACKGROUND: Identifying endogenous volatile organic compounds
(VOCs) as markers for different cancers currently requires time-consuming
procedures and specialized operators. OBJECTIVE: The objective of this study was to develop a rapid and
simple method for measuring VOCs at trace levels. METHODS: A simple vacuum ultraviolet photoionization mass
spectrometer (VUV-PIMS) was used to detect trace levels of dimethyl
trisulfide (DMTS), dimethyl sulfide (DMS), and 2-butanone, which correspond
to volatile biomarker candidates present in the exhaled breath of patients
with breast, liver, and lung cancers, respectively. The practicality of
measuring endogenous VOCs using VUV-PIMS was confirmed by detecting them in
cultured cell lines. RESULTS: The abovementioned VOCs were detected with high
sensitivity by VUV-PIMS. The limits of detection (LODs) for DMTS, DMS, and
2-butanone were 3.1, 3.9, and 23.2 pptv, respectively, under ambient
conditions, which surpass the sensitivity of nearly all other MS-based
techniques. Moreover, relatively high concentrations of 2-butanone and DMS
were observed in VOCs emitted from the A549 lung cancer cell line and the
HepG2 liver cancer cell line, respectively. CONCLUSIONS: Our
results show that VUV-PIMS may serve as a reliable method for real-time
measurement of endogenous volatile cancer biomarkers.
研究领域:
医学诊断
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分析化学
·
肿瘤学
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DOI
PubMed
高影响因子
Cancer Biomarkers
·
2021
·Vol. 30
·期 3
·331-342
·
IF 4.388
Yu Jiangqing,Du Fen,Yang Liping,Chen Ling,He Yuanxiang,Geng Ruijin,Wu Le,Xie Baogang
BACKGROUND: Histological subtypes of lung cancer are crucial for making treatment decisions. However, multi-subtype classifications including adenocarcinoma (AC), squamous cell carcinoma (SqCC) and small cell carcinoma (SCLC) were rare in the previous studies. This study aimed at identifying and screening potential serum biomarkers for the simultaneous classification of AC, SqCC and SCLC. PATIENTS AND METHODS: A total of 143 serum samples of AC, SqCC and SCLC were analyzed by 1HNMR and UPLC-MS/MS. The stepwise discriminant analysis (DA) and multilayer perceptron (MLP) were employed to screen the most efficient combinations of markers for classification. RESULTS: The results of non-targeted metabolomics analysis showed that the changes of metabolites of choline, lipid or amino acid might contribute to the classification of lung cancer subtypes. 17 metabolites in those pathways were further quantified by UPLC-MS/MS. DA screened out that serum xanthine, S-adenosyl methionine (SAM), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE) and squamous cell carcinoma antigen (SCC) contributed significantly to the classification of AC, SqCC and SCLC. The average accuracy of 92.3% and the area under the receiver operating characteristic curve of 0.97 would be achieved by MLP model when a combination of those five variables as input parameters. CONCLUSION: Our findings suggested that metabolomics was helpful in screening potential serum markers for lung cancer classification. The MLP model established can be used for the simultaneous diagnosis of AC, SqCC and SCLC with high accuracy, which is worthy of further study.
研究领域:
肿瘤学
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代谢组学
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医学
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DOI
PubMed
高影响因子
Cancer Biomarkers
·
2019
·Vol. 25
·期 2
·193-201
·
IF 2.859
Xu Jin-huan,Wang Yan,Xu Dong
Circular RNAs (circRNAs) have gained attention for their involvement in carcinogenesis, but its functional effects in breast cancer (BC) remains largely unclear. In this study, we aimed to explore the expressing pattern, clinical significance and potential function of a newly identified circRNA, hsa_circ_001569, in BC. RT-PCR was performed to detect the expression of hsa_circ_001569 in both BC tissues and cell lines. The associations between hsa_circ_001569 expression and clinicopathological features and prognosis in BC patients were statistically analyzed. Next, we investigated the effects of hsa_circ_001569 on the proliferation, apoptosis, migration and invasion in BC cells lines. The effects of abnormal hsa_circ_001569 expression on EMT pathway and PI3K/AKT pathway were determined using Western blot. We found that hsa_circ_001569 expression was significantly up-regulated in both BC tissues and cell lines. Overexpression of hsa_circ_001569 was associated with Lymph node metastasis, advanced clinical stage and shorter overall survival. Multivariate assay confirmed that hsa_circ_001569 expression was an independent prognostic factor for 5-year overall survival. Furthermore, functional investigations revealed that knockdown of hsa_circ_001569 significantly suppressed the growth and metastatic potentials of BC cells. Besides, molecular mechanistic study revealed that depression of hsa_circ_001569 impeded the activation of PI3K-AKT signaling in BC cells. Our results indicated that hsa_circ_001569 upregulation was associated with BC lymph-node metastasis, clinical stage, and poor prognosis. Hsa_circ_001569 might contribute to progression of BC by modulating PI3K-AKT pathway.
研究领域:
肿瘤学
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分子生物学
·
癌症研究
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DOI
PubMed
高影响因子
Cancer Biomarkers
·
2019
·Vol. 24
·期 4
·469-475
·
IF 2.859
Qin Yujing,Li Wenxue,Long Yingli,Zhan Zhijia
OBJECTIVE: This study aims to determine the correlation between p-cofilin expression and cisplatin resistance in patients with ovarian cancer, and also to investigate the role of p-cofilin in prognosis. PATIENTS AND METHODS: The ovarian cancer cell line A2780/DDP resistant to cisplatin was prepared. The cell resistance to cisplatin was measured via MTT assay. The cell invasion capacity was identified via Transwell assay. The mRNA expression and protein level was evaluated via semi-quantitative PCR and Western blot, respectively. The tumor tissues of patients with cisplatin-resistant ovarian cancer were collected. The relationship between prognosis and p-cofilin expression was analyzed. RESULTS: The growth rate of A2780 was similar to that of A2780/DDP. The sensitivity of A2780 to cisplatin was significantly higher than that of A2780/DDP (p< 0.01). The migration capacity of A2780/DDP was significantly increased compared to that of A2780 (p< 0.01), indicating that the cisplatin-resistant cell lines were successfully constructed. Both CFL1 mRNA level and p-cofilin level in A2780/DDP was significantly higher than that in A2780 (p< 0.01). The p-cofilin level in cancer tissues in patients with cisplatin-resistant ovarian cancer was significantly higher than that in patients with cisplatin-sensitive ovarian cancer (p< 0.01). The cisplatin resistance was positively correlated with the p-cofilin expression level (r= 0.802, p= 0.023). The survival time of patients with normal or low level of p-cofilin was significantly longer than that of patients with high expression. CONCLUSION: The cisplatin resistance of ovarian cancer is closely related to the expression level of p-cofilin, which affects the prognosis of patients with ovarian cancer.
研究领域:
肿瘤学
·
分子生物学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
Cancer Biomarkers
·
2018
·Vol. 23
·期 4
·603-613
·
IF 2.392
Liu Zhi-Biao,Tang Chen,Jin Xin,Liu Shou-Hou,Pi Wen
BACKGROUND: Small nucleolar RNA host gene 12 (SNHG12) has been shown to be a long noncoding RNA (lncRNA) that facilitates the progression of a number of malignancies. However, the expression pattern and biological function of SNHG12 in nasopharyngeal carcinoma (NPC) have not been investigated. OBJECTIVE: The aim of our study is to investigate the expression, clinical significance and function of SNHG12 in NPC. METHODS: RT-PCR was used to detect the expression of SNHG12 in NPC cell lines and primary tumor tissues. The correlation of SNHG12 with clinicopathological features and patient prognosis was analyzed. The biologic functions of SNHG12 in NPC were explored by MTT assay, colony formation assay, wound healing assays, transwell assay and flow cytometric analysis in vitro. The expression of EMT markers and Notch signal pathway markers were determined by western blotting. RESULTS: The expression levels of SNHG12 were up-regulated in both NPC tissues and cell lines. High SNHG12 expression was significantly associated with clinical stage, grade and poor prognosis. Multivariate analysis demonstrated that high lncRNA SNHG12 expression was an independent poor prognostic factor for NPC patients. Functionally, knockdown of SNHG12 suppressed NPC cells proliferation, migration and invasion. Mechanistic investigations showed that knockdown of SNHG12 suppressed the activation of EMT and Notch-1 signal pathway. CONCLUSIONS: Our data suggest that SNHG12 promotes the progression of NPC and is a potential therapeutic target for NPC intervention.
研究领域:
肿瘤学
·
分子生物学
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2026
·
IF 6.7
Fang Ruiqi,Chen JiaJia,Xu Yan,Yan Lishi,Ju Xin,Li Liangzhi
Biomass polysaccharides are explored
as inexpensive raw materials
for preparing value-added products, such as mannan oligosaccharides
(MOS). However, the varied composition of MOS and the underlying mechanism
have not been studied thoroughly yet. Here a β-mannanase from Paenibacillus sp. was engineered for hydrolyzing
biomass polysaccharides and producing prebiotic MOS. Structural analysis
and alanine scanning revealed two hot spots, Q66 and T88, in the carbohydrate
binding module (CBM). The mutant T88Y demonstrated a 65% activity
improvement and a 5-fold thermal stability enhancement at 70 ° C,
and Q66F produced 2.2 g/L of mannobiose and mannotriose from locust
bean gum, with a 46% activity increase. Moreover, Q66F generated a
2.0 molar ratio of mannobiose/mannotriose, and a reverse ratio of
0.4 was found in Q66R. The application of Q66R was further validated
in the fermentation of citrus peel waste. The results provided efficient
biocatalysts for producing prebiotic MOS from biomass polysaccharides
and deeper comprehension of altering product chain length of β-mannanases
from CBM.
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高影响因子
Polymers
·
2026
·Vol. 18
·期 7
·803
·
IF 5.8
Haibo Li,Lei Yue,Hongxing Xu,Yanhong Li,Yining Zhou,Rong Zhang,Kongjie Wang,Hongzhong Zhang,Shuai Luo,Bin Ren,Fei Chen,Yufei Liu
Coal-bed gas well production is too low to realize a highly efficient exploitation of the #8 coal seam in the Shanxi formation in the Nalin region. Based on the reservoir characteristics, the designed poly-aromatic-grafted silicon-quantum-dot-based desorption agent (PQS) has been developed. Then, the adsorption/desorption behavior of CBM on the coal surface under the influence of this active chemical has been studied, and the synergy effect with an anionic–nonionic surfactant to desorption of CBM has also been discussed. The results show that the developed poly-aromatic-grafted silicon quantum dot, with a median size of 4.9 nm and +5.6 mV of zeta potential in neutral condition, has a significant emission peak with 470 nm at the excitation of 380 nm and 150,000 mg/L of salinity resistance, which also generates a strong adsorption capacity on the coal surface. A promoting effect to desorption of CBM for PQS nanofluid is exhibited and the Langmuir pressure is obviously increased. However, when the PQS nanofluid is synergized with an anionic–nonionic surfactant, the desorption of CBM is further improved and the wettability of the coal surface is altered from 78.2° to 84.2°. The desorption rate for this compound system reached 65.3%. It can be found that combining the quantum size, π–π stacking, π–π conjugation, and the synergy effect between PQS nanofluid and surfactant fluid with the traditional intermolecular force has a stronger capacity for promoting desorption of CBM than the conventional desorption agent. This study provides guidance for the molecular design of the desorption agent for deep coal rock and the application of silicon quantum dots.
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高影响因子
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