Display of Lignin Peroxidase on the Surface of Bacillus subtilis.

Lignin peroxidase (LiP) has a good application prospect in lignin degradation, environmental treatment, straw feed, and other industries. However, its application is constrained by the high price and low stability of enzyme preparation. In this study, the Escherichia coli-Bacillus subtilis (E. coli-B. subtilis) shuttle expression vector pHS-cotG-lip was constructed and displayed on the surface of Bacillus subtilis spores. The analysis of enzymatic properties showed that the optimal catalytic temperature and pH of the immobilized LiP were 55 °C and 4.5, respectively. Compared with free LiP (42 °C and pH4.0), the optimal reaction temperature increased by 13 °C. After incubation at 70 °C for 1 h, its activity remained above 30%, while the free LiP completely lost its activity under the same conditions. Adding Mn2+, DL-lactic acid, and PEG-4000 increased the CotG-LiP enzyme activity to 313%, 146%, and 265%, respectively. The recyclability of spore display made the fusion protein CotG-LiP retain more than 50% enzyme activity after four cycles. The excellent recycling rate indicated that LiP displayed on the spore surface had a good application prospect in sewage treatment and other fields, and also provided a reference for the rapid and low-cost immobilized production of enzyme preparations.

Preparing the pure lignin peroxidase and exploring the effects of chemicals on the activity.

Heterogous expression of lignin peroxidase (LiP) from Phanerochaete chrysosporium was performed in by E. coli prokaryotic expression system, and pure LiP was prepared by washing, refolding, and purification. The enzyme activity was measured by the resveratrol oxidation method. The effects of different chemicals on LiP activity were explored by adding different kinds of metal ions, acids/phenols, and surfactants. The optimal pH and temperature are 4.2 and 40 °C. The single-factor screening experiment showed that adding 1 mM Mn2+, 0.1 mM DL-lactic acid, and 2% PEG-4000 had the best promotion effect on the enzyme activity of recombinant LiP, which was 160.61%, 188.46%, and 247.83%, respectively. Further, the synergistic addition of Mn2+ and PEG-4000 achieved the best enzyme activity promotion effect of 277.51%. In addition, the addition of DL-lactic acid alone could promote LiP activity. However, the co-addition of lactic acid with Mn2+ and PEG-4000 contributed only 247.87%, which indicated that the addition of DL-lactic acid had an inhibitory effect when applied synergistically. For the first time, it was found that PEG-4000 increased LiP enzyme activity obviously and had a synergistic effect with Mn2+, serving as a reference for LiP in studies and applications pertaining to lignin breakdown.

Straw lignin degradation by lignin peroxidase from Irpex lacteus cooperated with enzymes and small molecules.

OBJECTIVES: Maximizing the utility value of enzymes was achieved by exploring the effects of small molecules on the efficiency of lignin degradation by lignin peroxidase. METHODS: Using wheat straw as raw material and taking lignin degradation rate as index, it was found that laccase, glucose oxidase, malonic acid, citric acid, ZnSO4, CaCl2 could promote the lignin degradation by the lignin peroxidase from Irpex lacteus, respectively. Moreover, glucose oxidase, malonic acid and CaCl2 had obvious synergy effects on lignin degradation by the lignin peroxidase. RESULTS: The optimal conditions of lignin degradation were obtained by response surface experiment: 4% glucose oxidase, 0.74% malonic acid and 3.29% CaCl2 were added for synergistic degradation at 37 â„ƒ with 50% of water content. After 72 h quickly enzymatic hydrolysis, the degradation rate of lignin was 45.84%. CONCLUSIONS: A new green and efficient method for lignin removal from straw was obtained, which provided a reference for the efficient utilization of straw and lignin peroxidase.

Synergistic Degradation of Maize Straw Lignin by Manganese Peroxidase from Irpex lacteus.

Lignocellulose in maize straw includes cellulose, hemicellulose, and lignin, and the degradation of lignocellulose is a complex process in which multiple enzymes are jointly involved. In exploring the co-degradation of a certain substrate by multiple enzymes, different enzymes are combined freely for the achievement of the effective synergism. Additionally, some organic acids and small molecule aromatic compounds can also increase the enzymatic activity of lignin enzymes and improve the degradation rate of lignin. In this study, manganese peroxidase (MnP) from Irpex lacteus (I. lacteus) was heterologously expressed in food-grade Schizosaccharomyces pombe (S. pombe). The multiple enzymes co-fermentation conditions were initially screened by orthogonal tests: 0.5% CaCl2, 1% 10,000 U/g Laccase (Lac), 0.3% MnSO4, and 0.4% glucose oxidase (GOD). It was showed that the lignin degradation rate could reach 65.85% after 3 days of synergistic degradation with the addition of 0.02% Tween-80, 0.5 mM oxalic acid. This indicates that oxalic acid has a promoting effect on the activity of MnP, and the promoting effect is more significant when Tween-80 is complexed with oxalic acid.

Decreased MT1-MMP in gastric cancer suppressed cell migration and invasion via regulating MMPs and EMT.

Membrane type 1-matrix metalloproteinase (MT1-MMP) has been identified to play a significant role in several types of cancers, but little is known about the significance of MT1-MMP in gastric cancer patients. The purpose of this study is to investigate the involvement of MT1-MMP in tumor progression of gastric cancer. MT1-MMP expression levels were examined in gastric cancer tissues and cells, and normal gastric tissues and cells. The effects and molecular mechanisms of MT1-MMP expression on cell proliferation, migration, and invasion were also explored. In our results, MT1-MMP messenger RNA (mRNA) and protein expression levels were significantly increased in gastric cancer tissue. Moreover, the overexpression of MT1-MMP was positively associated with the status of clinical stage and lymph node metastasis through real-time PCR. Furthermore, knocking down MT1-MMP expression significantly suppressed the cell migration and invasion in vitro and regulated the expression of MMPs and epithelial-mesenchymal transition (EMT)-associated genes. In conclusions, our study demonstrates that MT1-MMP was overexpressed in gastric cancer tissue, and reduced expression of MT1-MMP suppressed cell migration, invasion, and through regulating the expression of MMPs and the process of EMT in gastric cancer.

Meta-analysis of laparoscopic versus open colorectal surgery within fast-track perioperative care.

BACKGROUND: Both laparoscopic surgery and fast-track perioperative care have demonstrated advantages in patients undergoing elective colorectal resections. It is unclear whether there is an additive effect by combining these 2 procedures. OBJECTIVE: The study aimed to conduct a meta-analysis of the randomized evidence to compare laparoscopic with open colorectal surgery within fast-track perioperative care. DATA SOURCES: PubMed, MEDLINE, Embase, and the Cochrane library databases were electronically searched (January 1985 to August 2011). STUDY SELECTION: Randomized clinical trials compared laparoscopic with open colorectal resections within fast-track programs. INTERVENTION: Investigators independently reviewed articles, extracted data, and assessed study quality according to standardized criteria. MAIN OUTCOME MEASURES: The main outcomes measured were postoperative hospital stay, overall hospital stay, readmission rate, morbidity, and mortality. RESULTS: Three trials were considered suitable for meta-analysis. A total of 171 patients underwent laparoscopic surgery, and 142 had open surgery. Meta-analysis showed that laparoscopic colorectal surgery had shorter postoperative hospital stays (weighted mean difference -1.06; 95% CI, -2.06 to -0.06, z = 2.08, p = 0.04) and shorter overall hospital stays (weighted mean difference -2.04; 95% CI, -3.50 to -0.58, z = 2.74, p < 0.01). No significant differences were noted for readmission rate (OR 0.54; 95% CI, 0.26-1.12, z = 1.65, p = 0.10), morbidity (OR 0.68; 95% CI, 0.42-1.10, z =1.58, p = 0.11), and mortality (OR 0.33; 95% CI, 0.09-1.18, z =1.70, p = 0.09). No publication bias and no significant heterogeneity were noted. LIMITATIONS: This study was limited because of its small sample size. CONCLUSIONS: Laparoscopic colorectal surgery has shorter postoperative hospital stays and overall hospital stays than open surgery within fast-track perioperative care. There is no significant difference with respect to readmission rate, morbidity, and mortality. Because the number of patients included in the present trials was small, further studies should be undertaken to confirm these findings.