Effect of nitrogen sources on oxidoreductive enzymes and ethanol production during D-xylose fermentation by Candida shehatae

1992 ◽  
Vol 38 (3) ◽  
pp. 258-260 ◽  
Author(s):  
Sanjay Palnitkar ◽  
Anil Lachke

The effect on D-xylose utilization and the corresponding xylitol and ethanol production by Candida shehatae (ATCC 22984) were examined with different nitrogen sources. These included organic (urea, asparagine, and peptone) and inorganic (ammonium chloride, ammonium nitrate, ammonium sulphate, and potassium nitrate) sources. Candida shehatae did not grow on potassium nitrate. Improved ethanol production (Y(p/s), yield coefficient (grams product/grams substrate), 0.34) was observed when organic nitrogen sources were used. Correspondingly, the xylitol production was also higher with organic sources. Ammonium sulphate showed the highest ethanol:xylitol ratio (11.0) among all the nitrogen sources tested. The ratio of NADH- to NADPH-linked D-xylose reductase (EC 1.1.1.21) appeared to be rate limiting during ethanologenesis of D-xylose. The levels of xylitol dehydrogenase (EC 1.1.1.9) were also elevated in the presence of organic nitrogen sources. These results may be useful in the optimization of alcohol production by C. shehatae during continuous fermentation of D-xylose. Key words: xylose fermentation, Candida shehatae, nitrogen source, oxidoreductive enzymes.

2020 ◽  
Author(s):  
Roksolana Vasylyshyn ◽  
Olena Kurylenko ◽  
Justyna Ruchala ◽  
Nadiya Shevchuk ◽  
Neringa Kuliesiene ◽  
...  

Abstract Background Xylose transport is one of the bottlenecks in the conversion of lignocellulosic biomass to ethanol. Xylose consumption by the wild-type strains of xylose-utilizing yeasts occurs once glucose is depleted resulting in a long fermentation process and overall slow and incomplete conversion of sugars liberated from lignocellulosic hydrolysates. Therefore, the engineering of endogenous transporters for the facilitation of glucose-xylose co-consumption is an important prerequisite for efficient ethanol production from lignocellulosic hydrolysates. Results In this study, several engineering approaches formerly used for the low-affinity glucose transporters in Saccharomyces cerevisiae , were successfully applied for earlier identified transporter Hxt1 in Ogataea polymorpha to improve xylose consumption (engineering involved asparagine substitution to alanine at position 358 and replacement of N-terminal lysine residues predicted to be the target of ubiquitination for arginine residues). Moreover, the modified versions of S. cerevisiae Hxt7 and Gal2 transporters also led to improved xylose fermentation when expressed in O. polymorpha . Conclusions The O. polymorpha strains with modified Hxt1 were characterized by simultaneous utilization of both glucose and xylose, in contrast to the wild-type and parental strain with elevated ethanol production from xylose. When the engineered Hxt1 transporter was introduced into constructed earlier advanced ethanol producer form xylose, the resulted strain showed further increase in ethanol accumulation during xylose fermentation. The overexpression of heterologous S. cerevisiae Gal2 had a less profound positive effects on sugars uptake rate, while overexpression of Hxt7 revealed the least impact on sugars consumption.


2020 ◽  
Author(s):  
Roksolana Vasylyshyn ◽  
Olena Kurylenko ◽  
Justyna Ruchala ◽  
Nadiya Shevchuk ◽  
Neringa Kuliesiene ◽  
...  

Abstract Background Xylose transport is one of the bottlenecks in the conversion of lignocellulosic biomass to ethanol. Xylose consumption by the wild-type strains of xylose-utilizing yeasts occurs once glucose is depleted resulting in a long fermentation process and overall slow and incomplete conversion of sugars liberated from lignocellulosic hydrolysates. Therefore, the engineering of endogenous transporters for the facilitation of glucose-xylose co-consumption is an important prerequisite for efficient ethanol production from lignocellulosic hydrolysates. Results In this study, several engineering approaches formerly used for the low-affinity glucose transporters in Saccharomyces cerevisiae , were successfully applied for earlier identified transporter Hxt1 in Ogataea polymorpha to improve xylose consumption (engineering involved asparagine substitution to alanine at position 358 and replacement of N-terminal lysine residues predicted to be the target of ubiquitination for arginine residues). Moreover, the modified versions of S. cerevisiae Hxt7 and Gal2 transporters also led to improved xylose fermentation when expressed in O. polymorpha . Conclusions The O. polymorpha strains with modified Hxt1 were characterized by simultaneous utilization of both glucose and xylose, in contrast to the wild-type and parental strain with elevated ethanol production from xylose. When the engineered Hxt1 transporter was introduced into constructed earlier advanced ethanol producer form xylose, the resulted strain showed further increase in ethanol accumulation during xylose fermentation. The overexpression of heterologous S. cerevisiae Gal2 had a less profound positive effects on sugars uptake rate, while overexpression of Hxt7 revealed the least impact on sugars consumption.


2018 ◽  
Vol 11 (1) ◽  
pp. 1-7
Author(s):  
C.E. Oshoma ◽  
Henrietta O. Obueh

Formic acid is one of the major inhibitory compounds present in hydrolysates derived from lignocellulosic materials, the presence of which can significantly hamper the efficiency of converting available sugars in the hydrolysates to bioethanol. The influence of different nitrogen sources supplemented media on bioethanol fermentation with Saccharomyces spp in the presence of formic acid was studied. The addition of ammonium sulphate and urea significantly increased cell number, glucose utilization, ethanol and glycerol production when compared with control media. It was observed that supplementation with nitrogen sources contributed assimilable nitrogen for the yeast strains growth when stressed with formic acid. The best concentration of nitrogen to be utilized by yeast was found to be 0.80 gN/L. The maximum cell numbers were 8.17± 0.23x 107 and 8.68± 0.16x 107 cells/mL for ammonium sulphate medium while control had the least of 4.00± 0.20x 107 and 4.97± 0.08x 107 cells/mL for S. cerevisiae NCYC2592 and S. arboricolus 2.3319 respectively. Ethanol accumulation increased with the consumption of glucose. The maximum ethanol production were 20.49 ± 1.24 and 19.74± 0.89 g/L using ammonium sulphate for S. cerevisiae NCYC2592 and S. arboricolus 2.3319 respectively. There was significant difference in ethanol production when the nitrogen sources were compared with the control medium (p< 0.05). The highest glycerol produced were 2.21 ± 0.04 and 3.11± 0.05 g/L using ammonium sulphate for S. cerevisiae NCYC2592 and S. arboricolus 2.3319 respectively. The conclusion was that yeast tolerance to formic acid and ethanol production could be achieved when fermentation medium is supplemented with nitrogen sources specifically ammonium sulphate.Keywords: Lignocellulose, inhibitors, Saccharomyces, fermentation, yeast, biofuel


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Morakot Krajang ◽  
Kwanruthai Malairuang ◽  
Jatuporn Sukna ◽  
Krongchan Rattanapradit ◽  
Saethawat Chamsart

Abstract Background A single-step ethanol production is the combination of raw cassava starch hydrolysis and fermentation. For the development of raw starch consolidated bioprocessing technologies, this research was to investigate the optimum conditions and technical procedures for the production of ethanol from raw cassava starch in a single step. It successfully resulted in high yields and productivities of all the experiments from the laboratory, the pilot, through the industrial scales. Yields of ethanol concentration are comparable with those in the commercial industries that use molasses and hydrolyzed starch as the raw materials. Results Before single-step ethanol production, studies of raw cassava starch hydrolysis by a granular starch hydrolyzing enzyme, StargenTM002, were carefully conducted. It successfully converted 80.19% (w/v) of raw cassava starch to glucose at a concentration of 176.41 g/L with a productivity at 2.45 g/L/h when it was pretreated at 60 °C for 1 h with 0.10% (v/w dry starch basis) of Distillase ASP before hydrolysis. The single-step ethanol production at 34 °C in a 5-L fermenter showed that Saccharomyces cerevisiae (Fali, active dry yeast) produced the maximum ethanol concentration, pmax at 81.86 g/L (10.37% v/v) with a yield coefficient, Yp/s of 0.43 g/g, a productivity or production rate, rp at 1.14 g/L/h and an efficiency, Ef of 75.29%. Scale-up experiments of the single-step ethanol production using this method, from the 5-L fermenter to the 200-L fermenter and further to the 3000-L industrial fermenter were successfully achieved with essentially good results. The values of pmax,Yp/s, rp, and Ef of the 200-L scale were at 80.85 g/L (10.25% v/v), 0.42 g/g, 1.12 g/L/h and 74.40%, respectively, and those of the 3000-L scale were at 70.74 g/L (8.97% v/v), 0.38 g/g, 0.98 g/L/h and 67.56%, respectively. Because of using raw starch, major by-products, i.e., glycerol, lactic acid, and acetic acid of all three scales were very low, in ranges of 0.940–1.140, 0.046–0.052, 0.000–0.059 (% w/v), respectively, where are less than those values in the industries. Conclusion The single-step ethanol production using the combination of raw cassava starch hydrolysis and fermentation of three fermentation scales in this study is practicable and feasible for the scale-up of industrial production of ethanol from raw starch.


2011 ◽  
Vol 80 (1) ◽  
pp. 149-167 ◽  
Author(s):  
G. Mulley ◽  
J. P. White ◽  
R. Karunakaran ◽  
J. Prell ◽  
A. Bourdes ◽  
...  

2015 ◽  
Vol 77 (31) ◽  
Author(s):  
Huszalina Hussin ◽  
Madihah Md Salleh ◽  
Chong Chun Siong ◽  
Muhammad Abu Naser ◽  
Suraini Abd- Aziz ◽  
...  

The recent study has demonstrated the effects of different nitrogen sources on vanillin production by Phanerochaete chrysosporium. Primary screening supported maximum biotransformation of ferulic acid (from lemongrass leaves hydrolysate) to vanillin by using ammonium chloride and yeast extract as inorganic and organic nitrogen source, respectively. With the 2-level factorial analysis, the optimum conditions of vanillin production from ferulic acid by P. chrysosporium was achieved at 0.192g/L with a molar yield of 24.5%.


Fermentation ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 313
Author(s):  
Paula Rojas ◽  
Daniel Lopez ◽  
Francisco Ibañez ◽  
Camila Urbina ◽  
Wendy Franco ◽  
...  

Organic nitrogen plays a significant role in the fermentation performance and production of esters and higher alcohols. This study assessed the use of yeast protein hydrolysate (YPH) as a nitrogen source for grape must fermentation. In this study, we prepared an enzymatic protein hydrolysate using yeasts recovered from a previous fermentation of wine. Three treatments were performed. DAP supplementation was used as a control, while two YPH treatments were used. Low (LDH) and high degrees of hydrolysis (HDH), 3.5% and 10%, respectively, were chosen. Gas chromatography and principal component analysis indicated a significant positive influence of YPH-supplementations on the production of esters and higher alcohols. Significantly high concentrations of 3-methyl-1-penthanol, isoamyl alcohol, isobutanol, and 2-phenylethanol were observed. Significant odorant activity was obtained for 3-methyl-1-pentanol and ethyl-2-hexenoate. The use of YPH as nitrogen supplementation is justified as a recycling yeasts technique by the increase in volatile compounds.


1962 ◽  
Vol 8 (1) ◽  
pp. 1-11 ◽  
Author(s):  
Jack McLachlan ◽  
P. R. Gorham

Microcystis aeruginosa Kütz. (strain NRC-1) grew equally well throughout the pH range 6.5 to 10 when provided with suitable media. Toxicity of tris(hydroxymethyl)aminomethane (TRIS) towards the alga was found to decrease as the pH decreased and could be correlated with the degree of ionization of the TRIS molecule. Other organic buffers examined were either toxic at all concentrations and pH values tested or promoted lysis. When TRIS was used as a buffer, higher concentrations of cesium chloride and potassium nitrate were tolerated without growth inhibition at pH 6.5 than at 7.5. In the presence of TRIS, Microcystis grew equally well with nitrate, ammonium, or urea as nitrogen sources. Eight out of 20 amino compounds examined served as nitrogen sources in TRIS-buffered medium, but growth was poorer than with nitrate nitrogen.


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