Hong Xu

Optimization of medium for one-step fermentation of inulin extract from Jerusalem artichoke tubers using Paenibacillus polymyxa ZJ-9 to produce R,R-2,3-butanediol.

The medium for one-step fermentation of raw inulin extract from Jerusalem artichoke tubers by Paenibacillus polymyxa ZJ-9 to produce R,R-2,3-butanediol (R,R-2,3-BD) was developed. Inulin, K(2)HPO(4) and NH(4)Cl were found to be the key factors in the fermentation according to the results obtained from the Plackett-Burman experimental design. The optimal concentration range of the three factors was examined by the steepest ascent path, and their optimal concentration was further investigated according to the Box-Behnken design and determined to be 77.14 g/L, 3.09 g/L and 0.93 g/L, respectively. Under the optimal conditions, the concentration of the obtained R,R-2,3-BD was 36.92 g/L, at more than 98% optical purity. Compared with other investigated carbon resources, fermentation of the raw inulin extract afforded the highest yield of R,R-2,3-BD. This process featured one-step fermentation of inulin without further hydrolyzing, which greatly decreased the raw material cost and thus facilitated its practical application.

Green and economical production of propionic acid by Propionibacterium freudenreichii CCTCC M207015 in plant fibrous-bed bioreactor.

Propionic acid production by Propionibacterium freudenreichii from molasses and waste propionibacterium cells was studied in plant fibrous-bed bioreactor (PFB). With non-treated molasses as carbon source, 12.69 ± 0.40 g l(-1) of propionic acid was attained at 120 h in free-cell fermentation, whereas the PFB fermentation yielded 41.22 ± 2.06 g l(-1) at 120 h and faster cells growth was observed. In order to optimize the fermentation outcomes, fed-batch fermentation was performed with hydrolyzed molasses in PFB, giving 91.89 ± 4.59 g l(-1) of propionic acid at 254 h. Further studies were carried out using hydrolyzed waste propionibacterium cells as substitute nitrogen source, resulting in a propionic acid concentration of 79.81 ± 3.99 g l(-1) at 302 h. The present study suggests that the low-cost molasses and waste propionibacterium cells can be utilized for the green and economical production of propionic acid by P. freudenreichii.

ε-Poly-L-lysine production by immobilized cells of Kitasatospora sp. MY 5-36 in repeated fed-batch cultures

The production of epsilon-PL by Kitasatospora sp. MY 5-36 through entrapment or adsorption on bagasse, synthetic sponge, macroporous silica gel, and loofah sponge was investigated in shake flask cultures, and immobilization on loofah sponge gave the highest epsilon-PL production. Repeated fed-batch cultures for epsilon-PL production were also carried out in a stirred bioreactor and final epsilon-PL concentrations and productivity of 34.11 g L(-1) and 9.34 g L(-1)d(-1), respectively were achieved by cells immobilize in loofah sponge. These values exceeded those for cultures with free cells (22.53 g L(-1) and 3.30 g L(-1)d(-1)). The immobilized cells were reused five times over a period of 526 h. These results suggest that the immobilization approach is promising for industrial applications.

L-Arabinose isomerase and its use for biotechnological production of rare sugars

L-Arabinose isomerase (AI), a key enzyme in the microbial pentose phosphate pathway, has been regarded as an important biological catalyst in rare sugar production. This enzyme could isomerize L-arabinose into L-ribulose, as well as D-galactose into D-tagatose. Both the two monosaccharides show excellent commercial values in food and pharmaceutical industries. With the identification of novel AI family members, some of them have exhibited remarkable potential in industrial applications. The biological production processes for D-tagatose and L-ribose (or L-ribulose) using AI have been developed and improved in recent years. Meanwhile, protein engineering techniques involving rational design has effectively enhanced the catalytic properties of various AIs. Moreover, the crystal structure of AI has been disclosed, which sheds light on the understanding of AI structure and catalytic mechanism at molecular levels. This article reports recent developments in (i) novel AI screening, (ii) AI-mediated rare sugar production processes, (iii) molecular modification of AI, and (iv) structural biology study of AI. Based on previous reports, an analysis of the future development has also been initiated.

Economical production of poly(γ-glutamic acid) using untreated cane molasses and monosodium glutamate waste liquor by Bacillus subtilis NX-2

The production of poly(γ-glutamic acid) by Bacillus subtilis NX-2 from cane molasses and monosodium glutamate waste liquor (MGWL) was studied for the first time in this work. When batch fermentation was carried out with untreated molasses, 33.6±0.37 g L(-1) PGA was obtained with a productivity of 0.46±0.006 g L(-1) h(-1). In order to minimize the substrate inhibition, fed-batch fermentation was performed with untreated or hydrolyzed molasses in 7.5 L bioreactor, giving 50.2±0.53 and 51.1±0.51 g L(-1) of PGA at 96 h, respectively. Further studies were carried out by using MGWL as another carbon source, resulting in a PGA concentration of 52.1±0.52 g L(-1) with a productivity of 0.54±0.003 g L(-1) h(-1). These results suggest that the low-cost cane molasses and MGWL can be used for the environmental-friendly and economical production of PGA by B. subtilis NX-2.

Propionic acid production in a plant fibrous-bed bioreactor with immobilized Propionibacterium freudenreichii CCTCC M207015

A plant fibrous-bed bioreactor (PFB) was constructed for propionic acid production. Sugar cane bagasse was applied to the PFB as immobilizing material. Starting at a concentration of 80g/L of glucose, Propionibacterium freudenreichii CCTCC M207015 produced 41.20±2.03g/L of propionic acid at 108h in the PFB. The value was 21.07% higher than that produced by free cell fermentation. Intermittent and constant fed-batch fermentations were performed in the PFB to optimize the fermentation results. The highest propionic acid concentration obtained from constant fed-batch fermentation was 136.23±6.77g/L, which is 1.40 times higher than the highest concentration (97.00g/L) previously reported. Scanning electron microscopy analysis showed that cells exhibited striking changes in morphology after PFB domestication. Compared with free cell fermentation, the fluxes of propionic acid synthesis and the pentose phosphate pathway in PFB fermentation increased by 84.65% and 227.62%, respectively. On the other hand, a decrease in succinic and acetic acid fluxes was also observed. The metabolic flux distributions of the two PFB fed-batch fermentation strategies also demonstrated that constant fed-batch fermentation is a more beneficial method for the immobilized production of propionic acid. The relevant key enzyme activities and metabolic flux variations of the batch cultures showed good consistency. These results suggest that the PFB was effective in high-concentration propionic acid production.

Enhanced poly(γ-glutamic acid) fermentation by Bacillus subtilis NX-2 immobilized in an aerobic plant fibrous-bed bioreactor

To enhance poly(γ-glutamic acid) (PGA) production, a novel aerobic plant fibrous-bed bioreactor (APFB) was constructed for immobilized fermentation. Based on the analysis of the kinetics of immobilized-cell fermentation using the APFB and conventional free-cell fermentation, immobilized-cell fermentation exhibited more efficient PGA production. Furthermore, repeated fed-batch cultures for PGA production were conducted to evaluate the stability of the APFB system. Average final PGA concentration and productivity of 71.21±0.83g/L and 1.246±0.008g/L/h were respectively achieved by cells immobilized in bagasse during APFB, which was reused eight times over a period of 457±18h. Analysis of the membrane phospholipids and the key enzyme activities indicated that APFB-adapted cells had better productivity than original cells. Thus, this study demonstrated the significant potential of the APFB culture system in future industrial applications.

Economical production of poly(ε-l-lysine) and poly(l-diaminopropionic acid) using cane molasses and hydrolysate of streptomyces cells by Streptomyces albulus PD-1.

Poly(ε-L-lysine) (ε-PL) and poly(L-diaminopropionic acid) (PDAP) co-production by Streptomyces albulus PD-1 from cane molasses and hydrolysate of strepyomyces cells (HSC) was investigated for the first time in this study. The optimal initial total sugar concentration of the cane molasses pretreated with sulfuric acid was determined to be 20 g L(-1), and HSC could substitute for yeast extract for ε-PL and PDAP co-production. When fed-batch fermentation was performed in 1t fermentor with pretreated cane molasses and HSC, 20.6 ± 0.5 g L(-1) of ε-PL and 5.2 ± 0.6 g L(-1) of PDAP were obtained. The amount of strepyomyces cells obtained in one fed-batch fermentation is sufficient to prepare the HSC to satisfy the demand of subsequent fermentations, thus the self-cycling of organic nitrogen source becomes available. These results suggest that the low-cost cane molasses and HSC can be used for the economical production of ε-PL and PDAP by S. albulus PD-1.

Conversion of agroindustrial residues for high poly(γ-glutamic acid) production by Bacillus subtilis NX-2 via solid-state fermentation.

Poly(γ-glutamic acid) (γ-PGA) production by Bacillus subtilis NX-2 was carried out through solid-state fermentation with dry mushroom residues (DMR) and monosodium glutamate production residues (MGPR; a substitute of glutamate) for the first time. Dry shiitake mushroom residue (DSMR) was found to be the most suitable solid substrate among these DMRs; the optimal DSMR-to-MGPR ratio was optimized as 12:8. To increase γ-PGA production, industrial waste glycerol was added as a carbon source supplement to the solid-state medium. As a result, γ-PGA production increased by 34.8%. The batch fermentation obtained an outcome of 115.6 g kg(-1) γ-PGA and 39.5×10(8) colony forming units g(-1) cells. Furthermore, a satisfactory yield of 107.7 g kg(-1) γ-PGA was achieved by compost experiment on a scale of 50 kg in open air, indicating that economically large-scale γ-PGA production was feasible. Therefore, this study provided a novel method to produce γ-PGA from abundant and low-cost agroindustrial residues.

Removal of Cr(III), Ni(II) and Cu(II) by poly(γ-glutamic acid) from Bacillus subtilis NX-2

Poly(γ-glutamic acid) (γ-PGA) derived from Bacillus subtilis NX-2 was investigated as a sorbent for heavy metal ions in batch adsorption experiments. The results showed that the heavy metal adsorption capacity of γ-PGA enhanced with the increase of pH, in the following order: Cr(III)>Cu(II)>Ni(II), within the pH range 3–5. The Langmuir sorption model effectively described the metal sorption of γ-PGA through the experiments of isotherm sorption, and it was deduced that the affinity of γ-PGA for metals was following the sequence: Cr(III)>Cu(II)>Ni(II). γ-PGA was also used to trap trace amounts of heavy metals from the electroplating wastewater, which were difficult to be entirely removed by the traditional hydroxide precipitation method. The results showed that Cr(III) and Ni(II) in the electroplating effluent decreased from 3.07 and 9.46 mg/l to 0.15 and 1.01 mg/l, respectively, and the treated solutions reached the effluent standard. Therefore, γ-PGA is satisfactory as a well biosorbent for the removal of heavy metals. The adsorption mechanism of γ-PGA binding heavy metals was also studied using HyperChem simulation and FT-IR.