Effect of an inorganic nitrogen source (NH4)2SO4 on the production of welan gum from Sphingomonas sp. mutant obtained through UV-ARTP compound mutagenesis.

As one of the most expensive extracellular polysaccharides, welan gum is widely used in biomedicine, food products, and petroleum because of its unique structure and excellent rheological properties. To reduce the cost of welan gum fermentation, together with (NH4)2SO4, which served as the sole nitrogen source, a high-welan-gum-producing mutant, B-8, screened through UV-ARTP compound mutagenesis was used. Under optimum conditions (C:N ratio 25:1, sucrose 50 g/L, (NH4)2SO4 4 g/L, and adding 8 mM NaCl at 32 h fermentation), the yield of welan gum and sucrose conversion were 18.86 g/L and 0.38 g/g, respectively, which were 98.95% and 137.50% higher than those achieved with the parent strain FM01, respectively. After the same treatment process, IN-welan (obtained with (NH4)2SO4) consumed less 95% ethanol, had higher molecular weight, and exhibited better rheological properties than ON-welan (obtained with beef extract). Transcriptome analysis revealed that (NH4)2SO4 could affect the synthetic pathway and monosaccharide content of welan gum by increasing bacterial chemotaxis and the availability of key intermediates. The fermentation performance of Sphingomonas sp. mutants could further be improved by providing several target genes to the mutants through metabolic engineering.

Effect of NaCl addition on the production of welan gum with the UV mutant of Sphingomonas sp.

Because of its excellent stability, non-toxicity, biodegradability and unique rheology, welan gum can be widely used in various fields, such as petroleum, biomedicine and food products. In this study, a high-yield mutant strain FM01-S09 was screened through two rounds of UV mutagenesis. Remarkably, the production of welan gum could be further increased by adding 4 mM NaCl at 32 h fermentation, reaching 30.12 ± 0.25 g/L (28.66% higher than no adding), and the NaCl-WG solution had stronger structural, impact resistance, and temperature resistance than H2O2-WG and WG solutions. Furthermore, the mechanism by which NaCl promotes welan gum synthesis was also investigated. It was found that cell membrane characteristics, intracellular microenvironment makeup, and key enzyme gene expression levels were significantly altered in different fermentation stages. Therefore, the addition of NaCl could effectively promote the growth and fermentation performance of Sphingomonas sp., providing a novel strategy for cost-effective welan gum production.

Production of welan gum from cane molasses by Sphingomonas sp. FM01.

As an extracellular exopolysaccharide, welan gum could serve as a suspending agent, stabilizer, emulsifier and thickener with characteristics that include bio-adhesion, non-toxicity, biocompatibility and biodegradability. In this study, Sphingomonas sp. FM01 was applied to effectively produce welan gum using cane molasses as the carbon source. After pretreatment with phosphoric acid and polyacrylamide, 60 g/L cane molasses combined with 6.0 g/L beef extract, 3 g/L KH2PO4, 0.5 g/L MgSO4, 15 mmol/L H2O2 and 4 mg/L niacin significantly improved the fermentation performance of Sphingomonas sp. FM01, increasing the production of welan gum to 37.65 g/L. Investigation of the rheological behavior of the welan gum obtained from the molasses-welan gum mixture showed it had an acceptable molecular weight and similar rheological properties and better viscosity stability compared to that obtained from sucrose, indicating that cane molasses could be explored as a suitable and inexpensive substrate for cost-effective welan gum production.

Pathway dissection, regulation, engineering and application: lessons learned from biobutanol production by solventogenic clostridia.

The global energy crisis and limited supply of petroleum fuels have rekindled the interest in utilizing a sustainable biomass to produce biofuel. Butanol, an advanced biofuel, is a superior renewable resource as it has a high energy content and is less hygroscopic than other candidates. At present, the biobutanol route, employing acetone-butanol-ethanol (ABE) fermentation in Clostridium species, is not economically competitive due to the high cost of feedstocks, low butanol titer, and product inhibition. Based on an analysis of the physiological characteristics of solventogenic clostridia, current advances that enhance ABE fermentation from strain improvement to product separation were systematically reviewed, focusing on: (1) elucidating the metabolic pathway and regulation mechanism of butanol synthesis; (2) enhancing cellular performance and robustness through metabolic engineering, and (3) optimizing the process of ABE fermentation. Finally, perspectives on engineering and exploiting clostridia as cell factories to efficiently produce various chemicals and materials are also discussed.

Effects of different expression systems on characterization of adenylate deaminase from Aspergillus oryzae.

Adenylate deaminase (AMPD) is an amino hydrolase that catalyzes the irreversible hydrolysis of adenosine monophosphate (AMP) to inosine monophosphate (IMP) and ammonia. In this study, the effect of different hosts on the enzymatic properties of AMPD from Aspergillus oryzae GX-08 was investigated and showed that Bacillus subtilis WB600 was more suitable for producing AMPD with a higher activity of 2540 U/mL. After purification, the optimal temperature and pH of recombinant AMPD were 55 °C and pH 6.0, respectively, and its activity was significantly enhanced by 10 mM Fe3+ with an increase of 236%. More importantly, the recombinant AMPD specifically and effectively catalyzed the conversion between AMP and IMP, in which 10 mL of crude AMPD achieved a conversion ratio of 76.4% after 40 min. Therefore, B. subtilis WB600 provides a potential platform for producing AMPD with excellent catalytic ability and catalytic specificity.