Ningjun Cao

Ethanol Production from Renewable Resources

Vast amounts of renewable biomass are available for conversion to liquid fuel, ethanol. In order to convert biomass to ethanol, the efficient utilization of both cellulose-derived and hemicellulose-derived carbohydrates is essential. Six-carbon sugars are readily utilized for this purpose. Pentoses, on the other hand, are more difficult to convert. Several metabolic factors limit the efficient utilization of pentoses (xylose and arabinose). Recent developments in the improvement of microbial cultures provide the versatility of conversion of both hexoses and pentoses to ethanol more efficiently. In addition, novel bioprocess technologies offer a promising prospective for the efficient conversion of biomass and recovery of ethanol.

Production of multifunctional organic acids from renewable resources.

Recently, the microbial production of multifunctional organic acid has received interest due to their increased use in the food industry and their potential as raw materials for the manufacture of biodegradable polymers. Certain species of microorganisms produce significant quantities of organic acids in high yields under specific cultivation conditions from biomass-derived carbohydrates. The accumulation of some acids, such as fumaric, malic and succinic acid, are believed to involve CO2-fixation which gives high yields of products. The application of special fermentation techniques and the methods for downstream processing of products are described. Techniques such as simultaneous fermentation and product recovery and downstream processing are likely to occupy an important role in the reduction of production costs. Finally, some aspects of process design and current industrial production processes are discussed.

Supercritical carbon dioxide explosion as a pretreatment for cellulose hydrolysis

SummaryCellulosic material Avicel was treated with supercritical carbon dioxide to increase the reactivity of cellulose, thereby to enhance the rate and the extent of cellulose hydrolysis. Upon an explosive release of the carbon dioxide pressure, the disruption of the cellulosic structure increases the accessible surface area of the cellulosic substrate to enzymatic hydrolysis. This explosion pretreatment enhances the rate of the Avicel hydrolysis as well as increases glucose yield by as much as 50%.

Dilute acid hemicellulose hydrolysates from corn cobs for xylitol production by yeast

Abstract Dilute acid hemicellulose hydrolysate, comprised mainly of xylose, was obtained from ground corn cobs after dilute hydrochloric acid (2%, wt) hydrolysis at 100°C for 2 h. Similar acid hydrolysate was also obtained after the corn cobs were treated with 10% ammonium hydroxide at 26°C for 24 h. Neutralized hydrolysates containing ca. 130 g/l xylose were used as the substrate for xylitol production by a xylitol-producing yeast, Candida sp. 11-2. Ammonia-treated hydrolysate was a better substrate for xylitol production. Xylose was consumed within 36 h with a specific xylitol productivity of 1.94 g/l·h and a xylitol weight yield of 0.57 g/g xylose utilized. In contrast, neutralized hemicellulose hydrolysate obtained without prior ammonia treatment required anion exchange resin treatment to be fermented by yeasts. The specific productivity and weight yield of xylitol from anion exchange resin-treated hydrolysate were comparable to that from an ammonia-treated sample.

Ethanol production from corn cob pretreated by the ammonia steeping process using genetically engineered yeast

SummaryA new and effective pretreatment process for biomass conversion involves the steeping of biomass in 2.9 M NH4OH. This resulted in the removing about 80–90% of the lignin along with almost all the acetate from cellulosic residues. Based on dry cellulose from corn cob, a high glucose yield of 92% was obtained after enzymatic saccharification of cellulose fraction. By using a genetically engineered, xylosefermenting Saccharomyces 1400(pLNH33) in the batch fermentation of a glucose-xylose mixture from corn cob, an ethanol concentration of 47 g/L was obtained within 36 h with 84% yield. In addition, an ethanol concentration of 45 g/L was obtained within 48 h with 86% yield using simultaneous saccharification-fermentation process.

Optimization of L-lactic acid production from glucose by Rhizopus oryzae ATCC 52311

The effect of nutrients on L(+)-lactic acid production from glucose was investigated using Rhizopus oryzae ATCC 52311. From the shake-flask experiments, the optimal medium composition was defined for improved lactic-acid production. In order to enhance lactic-acid production rate and product yield, controlled aeration in a bubble column was conducted under optimal conditions. Results showed a maximum lactic-acid production rate of 2.58 g/L/h was obtained with an initial glucose concentration of 94 g/L. Final lactic-acid concentration of 83 g/L was achieved after 32 h of fermentation with a weight of 0.88 g lactic acid/g glucose consumed.

Production of 2,3-butanediol from pretreated corn cob by Klebsiella oxytoca in the presence of fungal cellulase.

A simple and effective method of treatment of lignocellulosic material was used for the preparation of corn cob for the production of 2,3-butanediol byKlebsiella oxytoca ATCC 8724 in a simultaneous saccharification and fermentation process. During the treatment, lignin, and alkaline extractives were solubilized and separated from cellulose and hemicellulose fractions by dilute ammonia (10%) steeping. Hemicellulose was then hydrolyzed by dilute hydrochloric acid (1%, wJv) hydrolysis at 100°C at atmospheric pressure and separated from cellulose fraction. The remaining solid, with 90% of cellulose, was then used as the substrate. A butanediol concentration of 25 g/L and an ethanol concentration of 7 g/L were produced byK. oxytoca from 80 g/L of corn cob cellulose with a cellulase dosage of 8.5 IFPU/g corn cob cellulose after 72 h of SSF. With only dilute acid hydrolysis, a butanediol production rate of 0.21 g/L/h was obtained that is much lower than the case in which corn cob was treated with ammonia steeping prior to acid hydrolysis. The butanediol production rate for the latter was 0.36 g/L/h.

Production of Fumaric Acid by Immobilized Rhizopus Using Rotary Biofilm Contactor

Rotary biofilm contactor (RBC) is a reactor consisting of plastic discs that act as supports for micro-organisms. The discs are mounted on a horizontal shaft and placed in a medium-containing vessel. During nitrogen-rich growth phase, mycelia ofRhizopus oryzae ATCC 20344 grew on and around the discs and formed the “biofilm” of self-immobilized cells on the surface of the plastic discs. During the fermentation phase, the discs are slowly rotated, and the biofilms are exposed to the medium and the air space, alternately. With RBC, in the presence of CaCO3,Rhizopus biofilm consumes glucose and produces fumaric acid with a volumetric productivity of 3.78 g/L/h within 24 h. The volumetric productivity is about threefolds higher with RBC than with a stirred-tank fermenter with CaCO3. Furthermore, the duration of fermentation is one-third of the stirred-tank system. The immobilized biofilm is active for over a 2-wk period with repetitive use without loss of activity.

Cellulose hydrolysis using zinc chloride as a solvent and catalyst

Cellulose gel with < 10% of crystallinity was prepared by treatment of microcrystalline cellulose, Avicel, with zinc chloride solution at a ratio of zinc chloride to cellulose from 1.5 to 18 (w/w). The presence of zinc ions in the cellulose gels enhanced the rate of hydrolysis and glucose yield. The evidence obtained from X-ray diffraction, iodine absorption experiments; and Nuclear Magnetic Resonance spectra analysis suggested the presence of zinc-cellulose complex after Avicel was treated with zinc chloride. Zinc-cellulose complex was more susceptible to hydrolysis than amorphous cellulose. Under the experimental condition, cellulose gels with zinc ions were hyrolyzed to glucose with 95% theoretical yield and a concentration of 14% (w/v) by cellulases within 20 h. The same gel was hydrolyzed by acid to glucose with 91.5% yield and a concentration of 13.4% (w/v).

Ethanol production from xylose with the yeast Pichia stipitis and simultaneous product recovery by gas stripping using a gas-lift loop fermentor with attached side-arm (GLSA)

The bioconversion of xylose into ethanol with the yeast Pichia stipitis CBS 5773 is inhibited when 20 g/L of ethanol are present in the fermentation broth. In order to avoid this limitation, the fermentation was carried out with simultaneous recovery of product by CO(2) stripping. The fermentation was also improved by attaching a side-arm to the main body of a classical gas-lift loop fermentor. This side-arm increases the liquid circulation, mass transfer, and gas distribution, reducing the amount of oxygen in the inlet gas necessary to perform the fermentation of xylose under microaerobic conditions (K(L)a approximately 16 h(-1)). The continuous stripping of ethanol from the fermentation broth in this new bioreactor system allowed the consumption of higher xylose concentrations than using Erlenmeyer shaker flasks, improved significantly the process productivity and provided a clean ethanol solution by using an ice-cooled condenser system. Finally, a fed-batch fermentation was carried out with a K(L)a = 15.8 h(-1). Starting with 248.2 g of xylose, 237.6 g of xylose was consumed to produce 88.1 g of ethanol which represents 72.6% of the theoretical yield (47.2 g/L of ethanol was recovered in the condenser, while 9.6 g/L remained in the fermentation broth).