Dry Dilute Acid Research Articles

Improving xylose utilization and ethanol production from dry dilute acid pretreated corn stover by two-step and fed-batch fermentation

It is a great challenge to efficiently utilize xylose in fermentation with high solid loadings for the economically feasible production of lignocellulosic ethanol. However, 20% solid loading of dry dilute acid pretreated corn stover (DDAPCS) decreased 66.3% of the xylose consumption compared to yeast extract peptone dextrose and xylose (YPDX) medium, which was significantly larger than 16% solid loading (9.4%). To overcome those problems, several two-step processes for simultaneous saccharification and co-fermentation (SSCF) were developed by inoculating a water extract of 16% solid loading DDAPCS with microorganisms in the first fermentation step. During the first step, the relatively lower inhibitor concentrations (compared to 20% solid loading), decreased glucose concentration, and the absence of enzymatic hydrolysis residues (EHR) were beneficial for the use of xylose and enabled up to 75.6% of the xylose to be converted to ethanol. With an optimized fed-batch strategy of two-step process, the residual xylose concentration reduced by 55.6% with 20% solid loading, and the maximum ethanol concentration and yield increased by 30% and 16.6%, respectively.

Constructing xylose-assimilating pathways in Pediococcus acidilactici for high titer d-lactic acid fermentation from corn stover feedstock

Xylose-assimilating pathway was constructed in a d-lactic acid producing Pediococcus acidilactici strain and evolutionary adapted to yield a co-fermentation strain P. acidilactici ZY15 with 97.3g/L of d-lactic acid and xylose conversion of 92.6% obtained in the high solids content simultaneous saccharification and co-fermentation (SSCF) of dry dilute acid pretreated and biodetoxified corn stover feedstock. The heterologous genes encoding xylose isomerase (xylA) and xylulokinase (xylB) were screened and integrated into the P. acidilactici chromosome. The metabolic flux to acetic acid in phosphoketolase pathway was re-directed to pentose phosphate pathway by substituting the endogenous phosphoketolase gene (pkt) with the heterologous transketolase (tkt) and transaldolase (tal) genes. The xylose-assimilating ability of the newly constructed P. acidilactici strain was significantly improved by adaptive evolution. This study provided an important strain and process prototype for high titer d-lactic acid production from lignocellulose feedstock with efficient xylose assimilation.

Reduction of Reactor Corrosion by Eliminating Liquid-Phase Existence in Dry Dilute Acid Pretreatment of Corn Stover

Serious corrosion of reactor metal walls by aqueous dilute acid solution is one of the essential drawbacks of dilute acid pretreatment (DAP) in the biorefining of lignocellulose biomass. In this study, the corrosion of pretreatment reactor metal walls was significantly reduced by the dry dilute acid pretreatment (DDAP) of corn stover, a modified DAP, in which the aqueous dilute acid solution is completely absorbed by high solids corn stover feedstock. The iron ion release from the stainless-steel walls of the reactor was measured, and the corrosion degree was quantitatively characterized. The reactor corrosion was reduced by approximately 85% when the solid/liquid ratio increased from the conventional value of 1:10 to the high value of 1:0.5 at the same sulfuric acid concentration in the DDAP. The results provided the simple and practical solution to the key technical barrier of DAP and paved the way for large-scale application.

Lower pressure heating steam is practical for the distributed dry dilute sulfuric acid pretreatment

Most studies paid more attention to the pretreatment temperature and the resulted pretreatment efficiency, while ignored the heating media and their scalability to an industry scale. This study aimed to use a relative low pressure heating steam easily provided by steam boiler to meet the requirement of distributed dry dilute acid pretreatment. The results showed that the physical properties of the pretreated corn stover were maintained stable using the steam pressure varying from 1.5, 1.7, 1.9 to 2.1MPa. Enzymatic hydrolysis and high solids loading simultaneous saccharification and fermentation (SSF) results were also satisfying. CFD simulation indicated that the high injection velocity of the low pressure steam resulted in a high steam holdup and made the mixing time of steam and solid corn stover during pretreatment much shorter in comparison with the higher pressure steam. This study provides a design basis for the boiler requirement in distributed pretreatment concept.

Dual effect of soluble materials in pretreated lignocellulose on simultaneous saccharification and co-fermentation process for the bioethanol production

In this study, wash liquors isolated from ethylenediamine and dry dilute acid pretreated corn stover were used to evaluate the effect of soluble materials in pretreated biomass on simultaneous saccharification and co-fermentation (SSCF) for ethanol production, respectively. Both of the wash liquors had different impacts on enzymatic hydrolysis and fermentation. Enzymatic conversions of glucan and xylan monotonically decreased as wash liquor concentration increased. Whereas, with low wash liquor concentrations, xylose consumption rate, cell viability and ethanol yield were maximally stimulated in fermentation without nutrient supplementary. Soluble lignins were found as the key composition which promoted sugars utilization and cell viability without nutrient supplementary. The dual effects of soluble materials on enzymatic hydrolysis and fermentation resulted in the reduction of ethanol yield as soluble materials increased in SSCF.

Oxidative production of xylonic acid using xylose in distillation stillage of cellulosic ethanol fermentation broth by Gluconobacter oxydans

An oxidative production process of xylonic acid using xylose in distillation stillage of cellulosic ethanol fermentation broth was designed, experimentally investigated, and evaluated. Dry dilute acid pretreated and biodetoxified corn stover was simultaneously saccharified and fermented into 59.80g/L of ethanol (no xylose utilization). 65.39g/L of xylose was obtained in the distillation stillage without any concentrating step after ethanol was distillated. Then the xylose was completely converted into 66.42g/L of xylonic acid by Gluconobacter oxydans. The rigorous Aspen Plus modeling shows that the wastewater generation and energy consumption was significantly reduced comparing to the previous xylonic acid production process using xylose in pretreatment liquid. This study provided a practical process option for xylonic acid production from lignocellulose feedstock with significant reduction of wastewater and energy consumption.

Fermentative production of high titer citric acid from corn stover feedstock after dry dilute acid pretreatment and biodetoxification

The aim of this work is to study the citric acid fermentation by a robust strain Aspergillus niger SIIM M288 using corn stover feedstock after dry dilute sulfuric acid pretreatment and biodetoxification. Citric acid at 100.04g/L with the yield of 94.11% was obtained, which are comparable to the starch or sucrose based citric acid fermentation. No free wastewater was generated in the overall process from the pretreatment to citric acid fermentation. Abundant divalent metal ions as well as high titer of potassium, phosphate, and nitrogen were found in corn stover hydrolysate. Further addition of extra nutrients showed no impact on increasing citric acid formation except minimum nitrogen source was required. Various fermentation parameters were tested and only minimum regulation was required during the fermentation. This study provided a biorefining process for citric acid fermentation from lignocellulose feedstock with the maximum citric acid titer and yield.

Fermentative production of high titer gluconic and xylonic acids from corn stover feedstock by Gluconobacter oxydans and techno-economic analysis

High titer gluconic acid and xylonic acid were simultaneously fermented by Gluconobacter oxydans DSM 2003 using corn stover feedstock after dry dilute sulfuric acid pretreatment, biodetoxification and high solids content hydrolysis. Maximum sodium gluconate and xylonate were produced at the titer of 132.46g/L and 38.86g/L with the overall yield of 97.12% from glucose and 90.02% from xylose, respectively. The drawbacks of filamentous fungus Aspergillus niger including weak inhibitor tolerance, large pellet formation and no xylose utilization were solved by using the bacterium strain G. oxydans. The obtained sodium gluconate/xylonate product was highly competitive as cement retarder additive to the commercial product from corn feedstock. The techno-economic analysis (TEA) based on the Aspen Plus modeling was performed and the minimum sodium gluconate/xylonate product selling price (MGSP) was calculated as $0.404/kg. This study provided a practical and economic competitive process of lignocellulose utilization for production of value-added biobased chemicals.

In situ detoxification of dry dilute acid pretreated corn stover by co-culture of xylose-utilizing and inhibitor-tolerant Saccharomyces cerevisiae increases ethanol production

Co-culture of xylose-utilizing and inhibitor-tolerant Saccharomyces cerevisiae was developed for bioethanol production from undetoxified pretreated biomass in simultaneously saccharification and co-fermentation (SSCF) process. Glucose accumulation during late fermentation phase in SSCF using xylose-utilizing strain can be eliminated by the introduction of inhibitor-tolerant strain. Effect of different ratios of two strains was investigated and xylose-utilizing strain to inhibitor-tolerant strain ratio of 10:1 (w/w) showed the best xylose consumption and the highest ethanol yield. Inoculating of xylose-utilizing strain at the later stage of SSCF (24–48h) exhibited lower ethanol yield than inoculating at early stage (the beginning 0–12h), probably due to the reduced enzymatic efficiency caused by the unconsumed xylose and oligomeric sugars. Co-culture SSCF increased ethanol concentration by 21.2% and 41.0% comparing to SSCF using individual inhibitor-tolerant and xylose-utilizing strain (increased from 48.5 and 41.7g/L to 58.8g/L), respectively, which suggest this co-culture system was very promising.

Evaluation of soluble fraction and enzymatic residual fraction of dilute dry acid, ethylenediamine, and steam explosion pretreated corn stover on the enzymatic hydrolysis of cellulose

This study is aimed to examine the inhibition of soluble fraction (SF) and enzymatic residual fraction (ERF) in dry dilute acid (DDA), ethylenediamine (EDA) and steam explosion (SE) pretreated corn stover (CS) on the enzymatic digestibility of cellulose. SF of DDA, EDA and SE pretreated CS has high xylose, soluble lignin and xylo-oligomer content, respectively. SF of EDA pretreated CS leads to the highest inhibition, followed by SE and DDA pretreated CS. Inhibition of ERF of DDA and SE pretreated CS is higher than that of EDA pretreated CS. The inhibition degree (A0/A) of SF is 1.76 and 1.21 times to that of ERF for EDA and SE pretreated CS, respectively. The inhibition degree of ERF is 1.05 times to that of SF in DDA pretreated CS. The quantitative analysis shows that SF of EDA pretreated CS, SF and ERF of SE pretreated CS cause significant inhibition during enzymatic hydrolysis.

Acceleration of biodetoxification on dilute acid pretreated lignocellulose feedstock by aeration and the consequent ethanol fermentation evaluation.

BackgroundBiodetoxification by the fungus Amorphotheca resinae ZN1 provides an effective way of inhibitor removal from pretreated lignocellulose feedstock and has been applied in the process of ethanol, biolipids, and lactic acid production. However, the long-time used and the consumption of considerable xylose in the pretreated materials reduced the process efficiency. The improvements of biodetoxification should be made to enhance the production of biochemical from lignocellulosic materials.ResultsThis study reported an acceleration method of A. resinae ZN1-based biodetoxification on the corn stover (CS) feedstock pretreated using dry dilute acid pretreatment. Under proper aeration and well-mixing condition, the conversion rate of furfural, 5-hydroxymethylfurfural (HMF), acetic acid, and typical phenolic compounds were significantly accelerated by more than twofolds faster, which resulted in the reduction of biodetoxification time from 96 h in the conventional process to 36 h. Simultaneous saccharification and ethanol fermentation assay on accelerated biodetoxification of the dry dilute acid pretreated CS feedstock achieved the similar ethanol titer (48.56 g/L of 36 h’ accelerated biodetoxification vs. 50.00 g/L of 4 days’ conventional biodetoxification) and yield (58.10 vs. 59.63 %). Substrate priority of inhibitors to sugars by A. resinae ZN1 was discovered and considerable xylose was reserved in the accelerated biodetoxification. Cell growth of A. resinae fungus in liquid medium and on pretreated CS solids revealed that the enhanced aeration enhanced the biodetoxification rate rather than the cell growth rate. Accelerated inhibitor conversion might come from the increased supply of cofactors of nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate from the step of aldehyde inhibitors to the corresponding acids, instead of cell mass increase.ConclusionAccelerated biodetoxification reduced the period of biodetoxification and retained the xylose components in the pretreated CS, which provided a practical method on improving process efficiency for cellulosic ethanol production from severe pretreated lignocellulose feedstock.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0438-9) contains supplementary material, which is available to authorized users.

High titer gluconic acid fermentation by Aspergillus niger from dry dilute acid pretreated corn stover without detoxification

This study reported a high titer gluconic acid fermentation using dry dilute acid pretreated corn stover (DDAP) hydrolysate without detoxification. The selected fermenting strain Aspergillus niger SIIM M276 was capable of inhibitor degradation thus no detoxification on pretreated corn stover was required. Parameters of gluconic acid fermentation in corn stover hydrolysate were optimized in flasks and in fermentors to achieve 76.67g/L gluconic acid with overall yield of 94.91%. The sodium gluconate obtained from corn stover was used as additive for extending setting time of cement mortar and similar function was obtained with starch based sodium gluconate. This study provided the first high titer gluconic acid production from lignocellulosic feedstock with potential of industrial applications.

Engineering wild-type robust Pediococcus acidilactici strain for high titer l- and d-lactic acid production from corn stover feedstock

Pediococcus acidilactici TY112 producing l-lactic acid and P. acidilactici ZP26 producing d-lactic acid, were engineered from the wild-type P. acidilactici DQ2 by ldhD or ldh gene disruption, and the robustness of the wild-type strain to the inhibitors derived from lignocellulose pretreatment was maintained well. In simultaneous saccharification and fermentation (SSF), 77.66gL−1 of l-lactic acid and 76.76gL−1 of d-lactic acid were obtained at 25% (w/w) solids content of dry dilute acid pretreated and biodetoxified corn stover feedstock. l- and d-Lactic acid yield and productivity were highly dependent on the inhibitor removal extent due to the significant down-regulation on the expressions of ldh and ldhD encoding lactate dehydrogenase by inhibitor, especially syringaldehyde and vanillin at the low concentrations. This study provided a prototype of industrial process for high titer l- and d-lactic acid production from lignocellulose feedstock.

Long term storage of dilute acid pretreated corn stover feedstock and ethanol fermentability evaluation

This study reported a new solution of lignocellulose feedstock storage based on the distributed pretreatment concept. The dry dilute sulfuric acid pretreatment (DDAP) was conducted on corn stover feedstock, instead of ammonia fiber explosion pretreatment. Then the dry dilute acid pretreated corn stover was stored for three months during summer season with high temperature and humidity. No negative aspects were found on the physical property, composition, hydrolysis yield and ethanol fermentability of the long term stored pretreated corn stover, plus the additional merits including no chemicals recovery operation, anti-microbial contaminant environment from stronger acid and inhibitor contents, as well as the mild and slow hydrolysis in the storage. The new pretreatment method expanded the distributed pretreatment concept of feedstock storage with potential for practical application.

High titer l-lactic acid production from corn stover with minimum wastewater generation and techno-economic evaluation based on Aspen plus modeling

Technological potentials of l-lactic acid production from corn stover feedstock were investigated by experimental and techno-economic studies. An optimal performance with 104.5g/L in l-lactic acid titer and 71.5% in overall yield from cellulose in corn stover to l-lactic acid using an engineered Pediococcus acidilactici strain were obtained by overcoming several technical barriers. A rigorous Aspen plus model for l-lactic acid production starting from dry dilute acid pretreated and biodetoxified corn stover was developed. The techno-economic analysis shows that the minimum l-lactic acid selling price (MLSP) was $0.523perkg, which was close to that of the commercial l-lactic acid produced from starch feedstock, and 24% less expensive than that of ethanol from corn stover, even though the xylose utilization was not considered. The study provided a prototype of industrial application and an evaluation model for high titer l-lactic acid production from lignocellulose feedstock.

Simultaneous saccharification and co-fermentation of dry diluted acid pretreated corn stover at high dry matter loading: Overcoming the inhibitors by non-tolerant yeast

Dry dilute acid pretreatment (DDAP) is a promising method for lignocellulose bioconversion, although inhibitors generated during the pretreatment impede the fermentation severely. We developed the simultaneous saccharification and co-fermentation (SScF) of DDAP pretreated biomass at high solid loading using xylose fermenting Saccharomyces cerevisiae, SyBE005. Effect of temperature on SScF showed that ethanol yield at 34°C was 10.2% higher than that at 38°C. Ethanol concentration reached 29.5g/L at 15% (w/w) dry matter loading, while SScF almost ceased at the beginning at 25% (w/w) dry matter loading of DDAP pretreated corn stover. According to the effect of the diluted hydrolysate on the fermentation of strain SyBE005, a fed-batch mode was developed for the SScF of DDAP pretreated corn stover with 25% dry matter loading without detoxification, and 40.0g/L ethanol was achieved. In addition, high yeast inoculation improved xylose utilization and the final ethanol concentration reached 47.2g/L.

High ethanol fermentation performance of the dry dilute acid pretreated corn stover by an evolutionarily adapted Saccharomyces cerevisiae strain

Ethanol fermentation was investigated at the high solids content of the dry dilute sulfuric acid pretreated corn stover feedstock using an evolutionary adapted Saccharomyces cerevisiae DQ1 strain. The evolutionary adaptation was conducted by successively transferring the S. cerevisiae DQ1 cells into the inhibitors containing corn stover hydrolysate every 12h and finally a stable yeast strain was obtained after 65days’ continuous adaptation. The ethanol fermentation performance using the adapted strain was significantly improved with the high ethanol titer of 71.40g/L and the high yield of 80.34% in the simultaneous saccharification and fermentation (SSF) at 30% solids content. No wastewater was generated from pretreatment to fermentation steps. The results were compared with the published cellulosic ethanol fermentation cases, and the obvious advantages of the present work were demonstrated not only at the high ethanol titer and yield, but also the significant reduction of wastewater generation and potential cost reduction.

De-ashing treatment of corn stover improves the efficiencies of enzymatic hydrolysis and consequent ethanol fermentation

In this study, corn stover with different ash content was pretreated using dry dilute acid pretreatment method at high solids loading of 67% (w/w). The results indicate that the hydrolysis yield of corn stover is increased from 43.30% to 70.99%, and ethanol yield is increased from 51.74% to 73.52% when ash is removed from 9.60% to 4.98%. The pH measurement of corn stover slurry indicates that the decrease of pretreatment efficiency is due to the neutralization of sulfuric acid by alkaline compounds in the ash. The elemental analysis reveals that the ash has the similar composition with the farmland soil. This study demonstrates the importance of ash removal from lignocellulose feedstock under high solids content pretreatment.

Dry dilute acid pretreatment by co-currently feeding of corn stover feedstock and dilute acid solution without impregnation

Impregnation of lignocellulose materials with dilute acid solution is a routine operation in conventional dilute acid pretreatment. The dry dilute acid pretreatment (DDAP) at high solids content up to 70% is naturally considered to require longer impregnation time. In this study, a co-currently feeding operation of corn stover and dilute sulfuric acid solution without any impregnation was tested for DDAP. The DDAP pretreated corn stover without impregnation is found to be essentially no difference in pretreatment efficiency compared to those with impregnation in the helically agitated reactor. The yield from cellulose to ethanol in SSF again shows no obvious difference between the DDAP pretreated corn stover with and without impregnation. This study suggests that impregnation in DDAP was not necessary under the helical agitation mixing. The results provided a useful way of cost reduction and process simplification in pretreatment.

Helically agitated mixing in dry dilute acid pretreatment enhances the bioconversion of corn stover into ethanol.

BackgroundDry dilute acid pretreatment at extremely high solids loading of lignocellulose materials demonstrated promising advantages of no waste water generation, less sugar loss, and low steam consumption while maintaining high hydrolysis yield. However, the routine pretreatment reactor without mixing apparatus was found not suitable for dry pretreatment operation because of poor mixing and mass transfer. In this study, helically agitated mixing was introduced into the dry dilute acid pretreatment of corn stover and its effect on pretreatment efficiency, inhibitor generation, sugar production, and bioconversion efficiency through simultaneous saccharification and ethanol fermentation (SSF) were evaluated.ResultsThe overall cellulose conversion taking account of cellulose loss in pretreatment was used to evaluate the efficiency of pretreatment. The two-phase computational fluid dynamics (CFD) model on dry pretreatment was established and applied to analyze the mixing mechanism. The results showed that the pretreatment efficiency was significantly improved and the inhibitor generation was reduced by the helically agitated mixing, compared to the dry pretreatment without mixing: the ethanol titer and yield from cellulose in the SSF reached 56.20 g/L and 69.43% at the 30% solids loading and 15 FPU/DM cellulase dosage, respectively, corresponding to a 26.5% increase in ethanol titer and 17.2% increase in ethanol yield at the same fermentation conditions.ConclusionsThe advantage of helically agitated mixing may provide a prototype of dry dilute acid pretreatment processing for future commercial-scale production of cellulosic ethanol.