Characterization of a novel ribose-5-phosphate isomerase B from Curtobacterium flaccumfaciens ZXL1 for D-allose production.

Enzymatic preparation of rare sugars as an alternative to traditional sweeteners is an effective strategy to achieve a low-calorie healthy diet. Ribose-5-phosphate isomerase B (RpiB) is a key enzyme in the non-oxidative branch of the catalytic pentose phosphate pathway. Here, we investigated the potential of Curtobacterium flaccumfaciens ZXL1 (C. flaccumfaciens ZXL1) derived RpiB (CfRpiB) in D-allose preparation. The optimal reaction conditions for recombinant CfRpiB were found experimentally to be pH 7.0, 55 °C, and no metal ions. The kinetic parameters Km, kcat, and catalytic efficiency kcat/Km were 320 mM, 4769 s-1, and 14.9 mM-1 s-1 respectively. The conversion of D-allulose by purified enzyme (1 g L-1 ) to D-allose was 13% within 1 h. In addition, homology modeling and molecular docking were used to predict the active site residues: Asp13, Asp14, Cys72, Gly73, Thr74, Gly77, Asn106, and Lys144.

Bioactivity and health effects of garlic essential oil: A review.

Garlic (Allium sativum L.), the underground bulb of the Allium plant in the family Liliaceae, is a common and popular spice that has historically been used to prevent and treat many different diseases such as pain, deafness, diarrhea, tumors, and other healthy problems. Garlic essential oil contains a variety of organosulfur compounds, such as the most representative diallyl disulfides (DADS) and diallyl trisulfides (DATS), which have attracted great interest in medicine, food, and agriculture because of their rich biological activities. This paper reviews the research progress on the composition and bioactivities of garlic essential oil mixtures and the bioactivity of some typical monomeric sulfides in garlic essential oil. The active mechanisms of representative sulfides in garlic essential oil were analyzed, and the applications of garlic essential oil in functional food, food additives, and clinical treatment were discussed. Combined with the current research status, the limitations and development direction of garlic essential oil in the study of molecular mechanism were discussed, which is of great significance to the development of garlic essential oil as a natural and safe alternative medicine for treatment.

Efficient production of ε-poly-L-lysine from cassava bagasse hydrolysate used as carbon source by Streptomyces albulus US3-18.

The production of ε-poly-L-lysine (ε-PL) from cassava bagasse hydrolysate (CBH) by Streptomyces albulus US3-18 was investigated in this study. With 30 g/L glucose from CBH, 1.30 g/L ε-PL and 10.68 g/L biomass were obtained in shake flask fermentation. Interestingly, the two values were increased by 14.0% and 21.5%, respectively, compared to the control (1.14 g/L and 8.79 g/L). Simultaneously, the activities of four key enzymes of ε-PL synthesis during CBH fermentation were enhanced to varying degrees. In batch fermentation of 5-L bioreactor, 3.39 g/L ε-PL and 10.17 g/L DCW were harvested with 40 g/L glucose from CBH. The combination of fed-batch fermentation with two-stage pH strategy significantly increased ε-PL titer and biomass to 37.41 g/L and 41.0 g/L, respectively. Moreover, eleven volatile components were detected in CBH by GC-MS, and 6-pentyl-α-pyrone (6PP) was first identified as the most abundant volatile ingredient. The results in CBH fermentation demonstrated that S. albulus US3-18 exhibited high tolerance to these volatile byproducts. Using ICP-MS, the calcium concentration in CBH was determined as 195.0 mg/(kg hydrolyzate), and cobalt, copper, lead, chromium, mercury and arsenic were not detected. By adding 0.05 g/L CaCl2 to M3G medium, ε-PL yield was improved by 28.0%, indicating calcium was one of the factors for the enhanced ε-PL production. The study provides a reference for the efficient production of ε-PL from low-cost agricultural residues.

Efficient production of ε-poly-l-lysine using cassava starch and fish meal by Streptomyces albulus FQC-24.

ε-Poly-l-lysine (ε-PL) is used as a natural food preservative which consists of l-lysine units connected. However, due to the expensive culture medium, the production cost of ε-PL remains high. In this study, cheap raw materials cassava starch (CS) and fish meal (FM) were employed by S. albulus FQC-24 for ε-PL production. In the single factor experiment, the maximum ε-PL production reached 0.97 g/L at 60 g/L CS and 15 g/L FM. The results of screening experiments by Plackett-Burman design showed that three main components affecting ε-PL production were CS, FM, and (NH4)2SO4. And the standardized effects of CS, FM, and (NH4)2SO4 were 0.13, -0.22, and -0.2, respectively. The optimum fermentation medium developed by response surface methodology for ε-PL production contained (g/L) CS, 67.56; FM, 14.70 and (NH4)2SO4, 5.41. Under the optimum conditions, the ε-PL production was achieved 1.30 g/L, with 34.02% higher than that before optimization. Moreover, ε-PL productions of batch and fed-batch fermentation in a 7-L fermentor were improved to 2.13 and 17.17 g/L respectively, which increased by 0.64 and 12.2 times compared with the shake flask culture. The results indicated that FM and CS are promising substrates for the efficient production of ε-PL.

An enhanced ionic liquid-tolerant immobilized cellulase system via hydrogel microsphere for improving in situ saccharification of biomass.

A novel immobilized cellulase based on hydrogel microsphere was established to accommodate ionic liquid (IL)-involved in situ biomass saccharification. For immobilization, poly(N-isopropylacrylamide) (PNIPAM) was employed to immobilize cellulase, with immobilization yielded up to 95.6%. Besides, 1-ethyl-3-methyllimidazolium acetate ([EMIM]OAc) was found as an appropriate IL for in situ saccharification due to its relative enzymatic activity, which was as high as 167.5%. With utilization of immobilized cellulase in IL-pretreated in situ bagasse (BC) saccharification, relative enzymatic activity was 40.9%, which was higher than free cellulase under 5% (v/v) IL, and achieved 12.3%, which was found to be higher than free one in 25% (v/v) IL. Moreover, for estimating reusability, relative enzymatic activity on the 6th cycle retained over 85.2%. This novel immobilization system possessed a remarkable preservation to cellulase in hydrolysis contained interferences, e.g. IL, suggesting a noticeable practical potential in hydrolysis of cost-effective sustainable biomass materials.

Characterization of Ribose-5-Phosphate Isomerase B from Newly Isolated Strain Ochrobactrum sp. CSL1 Producing L-Rhamnulose from L-Rhamnose.

In this study, we attempted to find new and efficient microbial enzymes for producing rare sugars. A ribose-5-phosphate isomerase B (OsRpiB) was cloned, overexpressed, and preliminarily purified successfully from a newly screened Ochrobactrum sp. CSL1, which could catalyze the isomerization reaction of rare sugars. A study of its substrate specificity showed that the cloned isomerase (OsRpiB) could effectively catalyze the conversion of L-rhamnose to L-rhamnulose, which was unconventional for RpiB. The optimal reaction conditions (50°C, pH 8.0, and 1 mM Ca2+) were obtained to maximize the potential of OsRpiB in preparing L-rhamnulose. The catalytic properties of OsRpiB, including Km, kcat, and catalytic efficiency (kcat/Km), were determined as 43.47 mM, 129.4 sec-1, and 2.98 mM/sec. The highest conversion rate of L-rhamnose under the optimized conditions by OsRpiB could reach 26% after 4.5 h. To the best of our knowledge, this is the first successful attempt of the novel biotransformation of L-rhamnose to L-rhamnulose by OsRpiB biocatalysis.

Construction of an Efficient Mutant Strain of Trichosporonoides oedocephalis with HOG1 Gene Deletion for Production of Erythritol.

The mitogen-activated protein kinase HOG1 (high-osmolarity glycerol response pathway) plays a crucial role in the response of yeast to hyperosmotic shock. Trichosporonoides oedocephalis produces large amounts of polyols (,e.g., erythritol and glycerol) in a culture medium. However, the effects of HOG1 gene knockout and environmental stress on the production of these polyols have not yet been studied. In this study, a To-HOG1 null mutation was constructed in T. oedocephalis using the loxP-Kan-loxP/Cre system as replacement of the targeted genes, and the resultant mutants showed much smaller colonies than the wild-type controls. Interestingly, compared with the wild-type strains, the results of shake-flask culture showed that To-HOG1 null mutation increased erythritol production by 1.44-fold while decreasing glycerol production by 71.23%. In addition, this study investigated the effects of citric acid stress on the T. oedocephalis HOG1 null mutants and the wild-type strain. When the supplementation of citric acid in the fermentation medium was controlled at 0.3% (w/v), the concentration of erythritol produced from the wild-type and To-HOG1 knockout mutant strains improved by 18.21% and 21.65%, respectively.

Improved in situ saccharification of cellulose pretreated by dimethyl sulfoxide/ionic liquid using cellulase from a newly isolated Paenibacillus sp. LLZ1.

A cellulase producing strain was newly isolated from soil samples and identified as Paenibacillus sp. LLZ1. A novel aqueous-dimethyl sulfoxide (DMSO)/1-ethyl-3-methylimidazolium diethyl phosphate ([Emin]DEP)-cellulase system was designed and optimized. In the pretreatment, DMSO was found to be a low-cost substitute of up to 70% ionic liquid to enhance the cellulose dissolution. In the enzymatic saccharification, the optimum pH and temperature of the Paenibacillus sp. LLZ1 cellulase were identified as 6.0 and 40°C, respectively. Under the optimized reaction condition, the conversion of microcrystalline cellulose and bagasse cellulose increased by 39.3% and 37.6%, compared with unpretreated cellulose. Compared to current methods of saccharification, this new approach has several advantages including lower operating temperature, milder pH, and less usage of ionic liquid, indicating a marked progress in environmental friendly hydrolysis of biomass-based materials.

Fibrarecisin, a novel triterpenoid from Fibraurea recisa with antitumor activity.

Fibrarecisin (1), a novel triterpenoid, was isolated from chloroform extract of the stem bark of Fibraurea recisa Pierre. Its structure was elucidated by spectroscopic methods. Compound 1 exhibited significant antitumor activity in vitro against A-549 cancer cell at 10 microM by SRB and MTT methods.

[Application of element and metabolism balancing for the cultivation process with Streptomyces aureofaciens].

On the base of element and metablism balancing, the mathematical model of the cultivation process with Streptomyces aureofaciens was developed, and the unknown parameters in the model were estimated with the method of nonlinear optimization. Firstly the energetic coefficient of CTC biosynthesis was gained, which was 1.8 - 2.8 mol-ATP x C-mol(-1). The macroscopic reaction rates were predicted in the process and compared with the experimental values. The results show that the model can preferably describe the relationships between several macroscopic reaction rates in the process and can supervise the optimization of CTC fermentation process theoretically.