Bacillus megaterium as a companion strain in two-stage fermentation of vitamin C could secrete some active substances to spur growth of Gluconobacter oxydans to produce 2-KLG. In the fermenting system where Gluconobac...Bacillus megaterium as a companion strain in two-stage fermentation of vitamin C could secrete some active substances to spur growth of Gluconobacter oxydans to produce 2-KLG. In the fermenting system where Gluconobacter oxydans was combined with GB82-a mutated strain of B. megaterium by ion implantation, the amount of 2-KLG harvested was larger than that produced by the original B. megaterium BP52 being substituted for GB82. In this paper, we studied the effect of the active substances secreted by GB82 to enhance the capability of Gluconobacter oxydans to produce 2-KLG. The supernate of GB82 sampled at different cultivation times all had much more activity to spur Gluconobacter oxydans to yield 2-KLG than that of the original B. megaterium, which might be due to the genetic changes in the active components caused by ion implantation. Furthermore, the active substances of GB82's supernate would lose a part of its activity in extreme environments, which is typical of some proteins.展开更多
In the two-step vitamin C fermentation process, its precursor 2-keto-L-gulonic acid was synthesized from L-sorbose by mixed culture of Gluconobacter oxydans and Bacillus megaterium. The interaction between Gluconobact...In the two-step vitamin C fermentation process, its precursor 2-keto-L-gulonic acid was synthesized from L-sorbose by mixed culture of Gluconobacter oxydans and Bacillus megaterium. The interaction between Gluconobacter oxydans and Bacillus megaterium remains unclear and it is a challenge to mathematically model the mixed growth of these two strains. The Monod-type equations were previously proposed to describe the coupled growth of Gluconobacter oxydans and Bacillus megaterium. However, in this study, we modeled the interaction of these two strains in a macroscopic view by introducing the population theory. Taking account of the fact that the density or concentration of Gluconobacter oxydans or Bacillus megaterium was hardly to measure accurately in the mixed culture broth, the data of concentrations of the substrate and product were used to indirectly investigate the relation between these two strains. Three batch experiments were used to validate our model. And according to the values of identified parameters, the type of interaction between Gluconobacter oxydans and Bacillus megaterium was concluded to be predation, where Gluconobacter oxydans was predator, and Bacillus megaterium was prey.展开更多
Vinegars are commonly used as food condiments and preservatives. Apple cider vinegar (ACV) is also used in the Ayurvedic pharmaceutical industry because of its medicinal properties. Since specifically selected starter...Vinegars are commonly used as food condiments and preservatives. Apple cider vinegar (ACV) is also used in the Ayurvedic pharmaceutical industry because of its medicinal properties. Since specifically selected starter cultures for commercial vinegar production are not readily available, apple juice supplemented with sugar is commonly inoculated with a microbiologically undefined culture obtained from the previous batch of ACV. The present work focuses on the isolation of yeasts and acetic acid bacteria from ACV and the preparation of a starter culture. ACV was produced in a bench scale bioreactor using a traditional fermentation process wherein an acetic acid concentration of 3.8% was obtained after three weeks. Several acetic acid bacteria (AAB) were isolated from ACV using selective media. Microscopy revealed the cultures to be gram negative to gram variable short rods. The growth pattern of the isolates on differential media and biochemical tests suggested the presence of Acetobacter and Gluconobacter species. Ten potent isolates were selected for starter culture preparation. Two consortia were formulated with five AAB isolates in each along with a yeast isolate and used for ACV production, wherein an acetic acid concentration of 4.2% - 4.9% was obtained in 10 - 12 days. Thus, these two starter cultures with locally isolated AAB can be used for the commercial production of apple cider vinegar.展开更多
Amorphophallus konjac is rich in glucomannan,which can be hydrolyzed into glucose and mannose,thereby acting as an economic raw material for the acquisition of glucose and mannose.The total sugar yield was 91.2%when k...Amorphophallus konjac is rich in glucomannan,which can be hydrolyzed into glucose and mannose,thereby acting as an economic raw material for the acquisition of glucose and mannose.The total sugar yield was 91.2%when konjac powder was treated with 0.75%hydrochloric acid at 121℃for 1 h.Thus,dilute acid hydrolysates of konjac powder were used as a carbon source for obtaining value-added products.Here we showed that the microbial production of ethanol and mannonic acid was obtained by employing Candida shehatae(C.shehatae)and Gluconobacter oxydans(G.oxydans).Through a step-by-step bioprocess,glucose is the first selectively converted to ethanol by C.shehatae,which enables G.oxydans-mediated biocatalysis of mannose to mannonic acid.Finally,approximately 100 g ethanol and 340 g mannonic acid were produced starting from 1 kg refined konjac powder.The results demonstrated the feasibility of this bioconversion method for producing mannonic acid starting from crude hydrolysates of konjac powder.展开更多
2-keto-D-gluconic acid(2-KGA)is a key precursor for synthesising vitamin C and isovitamin C.However,phage contamination is as constant problem in industrial production of 2-KGA using Pseudomonas fluorescens.Gluconobac...2-keto-D-gluconic acid(2-KGA)is a key precursor for synthesising vitamin C and isovitamin C.However,phage contamination is as constant problem in industrial production of 2-KGA using Pseudomonas fluorescens.Gluconobacter holds promise for producing 2-KGA due to impressive resistance to hypertonicity and acids,and high utilisation of glucose.In this study,the 2-KGA synthesis pathway was regulated to enhance production of 2-KGA and reduce accumulation of the by-products 5-keto-D-gluconic acid(5-KGA)and D-gluconic acid(D-GA)in the 2-KGA producer Gluconobacter japonicus CGMCC 1.49.Knocking out the ga5dh-1 gene from a competitive pathway and overexpressing the ga2dh-A gene from the 2-KGA synthesis pathway via homologous recombination increased the titre of 2-KGA by 63.81%in shake flasks.Additionally,accumulation of 5-KGA was decreased by 63.52%with the resulting G.japonicas-Δga5dh-1-ga2dh-A strain.Using an intermittent fed-batch mode in a 3 L fermenter,2-KGA reached 235.3 g L^−1 with a 91.1%glucose conversion rate.Scaling up in a 15 L fermenter led to stable 2-KGA titre with productivity of 2.99 g L^−1 h^−1,11.99%higher than in the 3 L fermenter,and D-GA and 5-KGA by-products were completely converted to 2-KGA.展开更多
L-Sorbose is an essential intermediate for the industrial production of vitamin C(L-ascorbic acid).However,the formation of fructose and some unknown by-products significantly reduces the conversion ratio of D-sorbito...L-Sorbose is an essential intermediate for the industrial production of vitamin C(L-ascorbic acid).However,the formation of fructose and some unknown by-products significantly reduces the conversion ratio of D-sorbitol to L-sorbose.This study aimed to identify the key D-sorbitol dehydrogenases in Gluconobacter oxydans WSH-003 by gene knockout.Then,a total of 38 dehydrogenases were knocked out in G.oxydans WSH-003,and 23 dehydrogenase-deficient strains could increase L-sorbose production.G.oxydans-30,wherein a pyrroloquinoline quinone-dependent glucose dehydrogenase was deleted,showed a significant reduction of a by-product with the extension of fermentation time.In addition,the highest conversion ratio of 99.60%was achieved in G.oxydans MD-16,in which 16 different types of dehydrogenases were inactivated consecutively.Finally,the gene vhb encoding hemoglobin was introduced into the strain.The titer of L-sorbose was 298.61 g/L in a 5-L bioreactor.The results showed that the systematic engineering of dehydrogenase could significantly enhance the production of L-sorbose.展开更多
文摘Bacillus megaterium as a companion strain in two-stage fermentation of vitamin C could secrete some active substances to spur growth of Gluconobacter oxydans to produce 2-KLG. In the fermenting system where Gluconobacter oxydans was combined with GB82-a mutated strain of B. megaterium by ion implantation, the amount of 2-KLG harvested was larger than that produced by the original B. megaterium BP52 being substituted for GB82. In this paper, we studied the effect of the active substances secreted by GB82 to enhance the capability of Gluconobacter oxydans to produce 2-KLG. The supernate of GB82 sampled at different cultivation times all had much more activity to spur Gluconobacter oxydans to yield 2-KLG than that of the original B. megaterium, which might be due to the genetic changes in the active components caused by ion implantation. Furthermore, the active substances of GB82's supernate would lose a part of its activity in extreme environments, which is typical of some proteins.
基金the National Natural Science Foundation of China(No.60974068)
文摘In the two-step vitamin C fermentation process, its precursor 2-keto-L-gulonic acid was synthesized from L-sorbose by mixed culture of Gluconobacter oxydans and Bacillus megaterium. The interaction between Gluconobacter oxydans and Bacillus megaterium remains unclear and it is a challenge to mathematically model the mixed growth of these two strains. The Monod-type equations were previously proposed to describe the coupled growth of Gluconobacter oxydans and Bacillus megaterium. However, in this study, we modeled the interaction of these two strains in a macroscopic view by introducing the population theory. Taking account of the fact that the density or concentration of Gluconobacter oxydans or Bacillus megaterium was hardly to measure accurately in the mixed culture broth, the data of concentrations of the substrate and product were used to indirectly investigate the relation between these two strains. Three batch experiments were used to validate our model. And according to the values of identified parameters, the type of interaction between Gluconobacter oxydans and Bacillus megaterium was concluded to be predation, where Gluconobacter oxydans was predator, and Bacillus megaterium was prey.
文摘Vinegars are commonly used as food condiments and preservatives. Apple cider vinegar (ACV) is also used in the Ayurvedic pharmaceutical industry because of its medicinal properties. Since specifically selected starter cultures for commercial vinegar production are not readily available, apple juice supplemented with sugar is commonly inoculated with a microbiologically undefined culture obtained from the previous batch of ACV. The present work focuses on the isolation of yeasts and acetic acid bacteria from ACV and the preparation of a starter culture. ACV was produced in a bench scale bioreactor using a traditional fermentation process wherein an acetic acid concentration of 3.8% was obtained after three weeks. Several acetic acid bacteria (AAB) were isolated from ACV using selective media. Microscopy revealed the cultures to be gram negative to gram variable short rods. The growth pattern of the isolates on differential media and biochemical tests suggested the presence of Acetobacter and Gluconobacter species. Ten potent isolates were selected for starter culture preparation. Two consortia were formulated with five AAB isolates in each along with a yeast isolate and used for ACV production, wherein an acetic acid concentration of 4.2% - 4.9% was obtained in 10 - 12 days. Thus, these two starter cultures with locally isolated AAB can be used for the commercial production of apple cider vinegar.
基金the financial support from the National Natural Science Foundation of China(31901270)the Scientific Research Start-up Funds of Nanjing Forestry University,China(163030127).
文摘Amorphophallus konjac is rich in glucomannan,which can be hydrolyzed into glucose and mannose,thereby acting as an economic raw material for the acquisition of glucose and mannose.The total sugar yield was 91.2%when konjac powder was treated with 0.75%hydrochloric acid at 121℃for 1 h.Thus,dilute acid hydrolysates of konjac powder were used as a carbon source for obtaining value-added products.Here we showed that the microbial production of ethanol and mannonic acid was obtained by employing Candida shehatae(C.shehatae)and Gluconobacter oxydans(G.oxydans).Through a step-by-step bioprocess,glucose is the first selectively converted to ethanol by C.shehatae,which enables G.oxydans-mediated biocatalysis of mannose to mannonic acid.Finally,approximately 100 g ethanol and 340 g mannonic acid were produced starting from 1 kg refined konjac powder.The results demonstrated the feasibility of this bioconversion method for producing mannonic acid starting from crude hydrolysates of konjac powder.
基金the National Key Research and Development Program of China(2017YFC1600403)the National Natural Science Foundation of China(31830068,21822806)+2 种基金the Fundamental Research Funds for the Central Universities(JUSRP51701A)the National First-class Discipline Program of Light Industry Technology and Engineering(LITE2018-08)the Distinguished Professor Project of Jiangsu Province.
文摘2-keto-D-gluconic acid(2-KGA)is a key precursor for synthesising vitamin C and isovitamin C.However,phage contamination is as constant problem in industrial production of 2-KGA using Pseudomonas fluorescens.Gluconobacter holds promise for producing 2-KGA due to impressive resistance to hypertonicity and acids,and high utilisation of glucose.In this study,the 2-KGA synthesis pathway was regulated to enhance production of 2-KGA and reduce accumulation of the by-products 5-keto-D-gluconic acid(5-KGA)and D-gluconic acid(D-GA)in the 2-KGA producer Gluconobacter japonicus CGMCC 1.49.Knocking out the ga5dh-1 gene from a competitive pathway and overexpressing the ga2dh-A gene from the 2-KGA synthesis pathway via homologous recombination increased the titre of 2-KGA by 63.81%in shake flasks.Additionally,accumulation of 5-KGA was decreased by 63.52%with the resulting G.japonicas-Δga5dh-1-ga2dh-A strain.Using an intermittent fed-batch mode in a 3 L fermenter,2-KGA reached 235.3 g L^−1 with a 91.1%glucose conversion rate.Scaling up in a 15 L fermenter led to stable 2-KGA titre with productivity of 2.99 g L^−1 h^−1,11.99%higher than in the 3 L fermenter,and D-GA and 5-KGA by-products were completely converted to 2-KGA.
基金This work was supported by the National Natural Science Foundation of China(Key Program,31830068).
文摘L-Sorbose is an essential intermediate for the industrial production of vitamin C(L-ascorbic acid).However,the formation of fructose and some unknown by-products significantly reduces the conversion ratio of D-sorbitol to L-sorbose.This study aimed to identify the key D-sorbitol dehydrogenases in Gluconobacter oxydans WSH-003 by gene knockout.Then,a total of 38 dehydrogenases were knocked out in G.oxydans WSH-003,and 23 dehydrogenase-deficient strains could increase L-sorbose production.G.oxydans-30,wherein a pyrroloquinoline quinone-dependent glucose dehydrogenase was deleted,showed a significant reduction of a by-product with the extension of fermentation time.In addition,the highest conversion ratio of 99.60%was achieved in G.oxydans MD-16,in which 16 different types of dehydrogenases were inactivated consecutively.Finally,the gene vhb encoding hemoglobin was introduced into the strain.The titer of L-sorbose was 298.61 g/L in a 5-L bioreactor.The results showed that the systematic engineering of dehydrogenase could significantly enhance the production of L-sorbose.
