Superfine grinding of steam-exploded rice straw and its enzymatic hydrolysis

The combination of low severity steam explosion and superfine grinding has been studied with respect to side products generation and enzymatic hydrolysis efficiency. Chemical compositions, fiber characteristics and composed cells contents in the superfine ground product and the ground residue particles produced by superfine grinding were also studied. At the determined parameters using FJM-200 fluidized bed opposed jet mill, 78% superfine ground steam-exploded rice straw (SERS) products with the mean fiber length of 60 μm were obtained, the particles yield was 179% higher than that from the native rice straw (RS). Enzymatic hydrolysis, chemical composition, fiber characteristics and composed cells proportion of the superfine ground SERS product and the ground residue all show great differences. The difference in enzymatic hydrolysis and structural properties indicates that superfine grinding is a good way to fractionate SERS into easily bio-converted part and difficultly hydrolysed part.     

白腐菌及黑曲霉所产的纤维素复合酶对稻草秸秆的生物降解

研究了白腐菌及纤维素复合酶对稻草秸秆的协同生物降解。结果表明,利用黄孢原毛平革菌固态发酵稻草秸秆的过程中,LiP和MnP的最大活力可以达到28.3U/g和12.6U/g,同时,秸秆中的木质素能被有效降解,但纤维素、半纤维素降解率较低。添加黑曲霉所产的纤维素复合酶能有效地促进秸秆腐熟程度。在接入白腐菌培养10天后,每克稻草添加3 IU纤维素酶液并酶解48h可以使稻草秸秆中纤维素降解53.8%,半纤维素降解57.8%,木质素降解44.5%,干物质损失46.3%。此时细胞壁出现大范围破损,整个组织变得松散,秸秆完全腐熟。     

Production of levulinic acid from steam exploded rice straw via solid superacid, S2O8(2-)/ZrO2-SiO2-Sm2O3

In this study, solid superacid was employed to catalyze the decomposition of steam exploded rice straw (SERS) for the production of levulinic acid, a versatile platform chemical. The results revealed that solid superacid, S(2)O(8)(2-)/ZrO(2)-SiO(2)-Sm(2)O(3), could be used as a substitute for homogenous acid to catalyze the production of LA from SERS and LA yield increased with the addition of solid superacid. It was also found that steam explosion combined with superfine grinding of rice straw could effectively increase LA yield for reducing particle size of rice straw and enhancing the accessibility of cellulose. Under optimal conditions of 200°C, 10 min, 13.3% of solid superacid to pretreated rice straw, and 1:15 of solid-liquid ratio, LA yield of the superfine grinding SERS was 70% of the theoretical yield, which was equivalent with the homogeneous acid-catalyzed production of LA.     

Microbial Fermentation of Rice Straw: Nutritive Composition and In Vitro Digestibility of the Fermentation Products

Rice straw was fermented with Cellulomonas sp. and Alcaligenes faecalis. Microbial cells and undigested residue, as well as chemically treated (NaOH or NH₄OH) and untreated straws, were analyzed for nutrient composition and in vitro digestibility. In a typical fermentation, 75% of the rice straw substrate was digested, and 18.6% of the total substrate weight that disappeared was recovered as microbial protein. The microbial cell fraction was 37% protein and 5% crude fiber; the residue was 12% protein and 45% crude fiber. The microbial protein amino acid profile was similar to alfalfa, except for less cysteine. The microbial cells had more thiamine and less niacin than Torula yeast. In vitro digestibility of the microbial protein was 41.2 to 55%; that of cellulose was 52%.     

Enzymatic hydrolysis of cellulosic materials by Sclerotium rolfsii culture filtrate for sugar production.

The hydrolysis of purified celluloses (cotton, Avicel, Cellulose-123, Solka Floc SW40) and cellulosic wastes (rice straw, sugarcane bagasse, wood powders, paper factory effluents) by Sclerotium rolfsii CPC 142 culture filtrate was studied. Factors which effect saccharification such as pH, temperature, enzyme concentration, substrate concentration, produce inhibition, adsorption, and inactivation of enzyme and particle size were studied. Virtually no inhibition (less than 3%) of cellulose hydrolysis by the culture filtrate was observed by cellobiose and glucose up to 100 mg/mL. Filter paper degrading enzyme(s) (but neither carboxymethylcellulase nor beta-glucosidase) was adsorbed on cellulose. The n value in the S. rolfsii system was calculated to be 0.32 for Avicel P.H. 101 and 0.53 for alkali-treated (AT) rice straw indicating penetration of cellulase into AT rice straw. In batch experiments at 10% substrate level, solutions containing 6 to 7%, 3.8 to 4.7%, 4.0 to 5.1%, and 4.2 to 4.9% reducing sugars were produced in 24 to 48 from AT rice straw. AT bagasse, alkali - peracetic acid treated mesta wood and paper factory sedimented sludge effluent, respectively. The main constituent in the hydrolysate from cellulose was glucose with little or no cellobiose, probably due to the high cellobiase content in the culture filtrate.     

DiSC (direct saccharification of culms) process for bioethanol production from rice straw

A simple process (the direct-saccharification-of-culms (DiSC) process) to produce ethanol from rice straw culms, accumulating significant amounts of soft carbohydrates (SCs: glucose, fructose, sucrose, starch and β-1,3-1,4-glucan) was developed. This study focused on fully mature culms of cv. Leafstar, containing 69.2% (w/w of dried culms) hexoses from SCs and cellulose. Commercially-available wind-separation equipment successfully prepared a culm-rich fraction with a SC recovery of 83.1% (w/w) from rice straw flakes (54.1% of total weight of rice straw). The fraction was suspended in water (20%, w/w) for starch liquefaction, and the suspension was subjected to a simultaneous saccharification and fermentation with yeast, yielding 5.6% (w/v) ethanol (86% of the theoretical yield from whole hexoses in the fraction) after 24 h fermentation. Thus, the DiSC process produced highly-concentrated ethanol from rice straw in a one vat process without any harsh thermo-chemical pretreatments.     

Use of molecular-genetically bred Coprinus cinereus strains for an efficient isolation of cellulose from rice straw

Molecular-bred Coprinus cinereus monokaryotic strains with high lignin- and xylan-degrading activities were mixed-cultured at 27 degrees C in the liquid medium containing 0.5% (w/v) cut rice straw and 0.025% MnCl2. After 3 weeks, the culture supernatant was extensively treated with crude cellulase, showing the presence in it of 9.3% of the total cellulose of rice straw. When rice straw treated with 0.1 N NaOH or cultured with Ganoderma applanatum were used, the recoveries of the cellulose increased up to 29%. The same experiments were done by using a non-bred control strain, showing the recoveries of the cellulose from the treated or cultured rice straw to be 8%.     

Optimal conditions for pretreatment of rice straw with n-butylamine for enzymatic solubilization

Summary Three different pretreatment methods with n-butylamine (n-BA) were used to obtain fermentable sugars in a high yield from rice straw. The optimal conditions of each method were as follows: treating at boiling point for 1 h under refluxing in 10 w/w% n-BA with the weight ratio of n-BA to original rice straw more than 1.0, autoclaving at 120°C for 1 h in 1 w/w% n-BA with the weight ratio more than 0.1, and wetting for 2 h with the circulating condensate of the vapour evaporated from 2.5 w/w% n-BA with the weight ratio more than 0.8. Soaking rice straw with n-BA before the above pretreatments was not needed. For the circulation pretreatment, the overall cumulative yield of total sugars (70% of cellulose and hemicellulose in original rice straw) was best for both pretreatment and enzymatic solubilization steps, because there was no decomposition of monosaccharides during the pretreatment. Furthermore, the optimal degree of delignification for enzymatic solubilization of the pretreated rice straw was approximately 60% of lignin in the original.     

Preparation and characterization of cellulose nanocrystals from rice straw

Pure cellulose have been isolated from rice straw at 36% yield and hydrolyzed (64% H₂SO₄, 8.75 mL/g, 45 °C) for 30 and 45 min to cellulose nanocrystals (CNCs), i.e., CNC30 and CNC45, respectively. CNC45 was smaller (11.2 nm wide, 5.06 nm thick and 117 nm long) than CNC30 (30.7 nm wide, 5.95 nm thick and 270 nm long). Freeze-drying of diluted CNC suspensions showed both assembled into long fibrous structures: ultra-fine fibers (∼400 nm wide) from CNC45 and 1-2 μm wide broad ribbons interspersed with CNC clusters from CNC30. The self-assembled fibers from CNC30 and CNC45 were more highly crystalline (86.0% and 91.2%, respectively) and contained larger crystallites (7.36 nm and 8.33 nm, respectively) than rice straw cellulose (61.8%, 4.42 nm). These self-assembled fibers had essentially nonporous or macroporous structures with the CNCs well aligned along the fiber axis. Furthermore, the self-assembled ultra-fine fibers showed extraordinary structural stability, withstanding vigorous shaking and prolong stirring in water.     

Studies on untreated and urea-treated rice straw from three cultivation seasons: 2. Evaluation of straw quality through in vitro gas production and in sacco degradation measurements

Untreated and urea-treated straw and straw fractions of seven rice varieties from three cultivation seasons have been evaluated on their DM, OM loss and degradation characteristics from in sacco disappearance and in vitro gas production measurements. Drying temperatures from 45°C to 100°C did not seem to influence the degradability of urea-treated rice straw, whereas urea-treated straw dried at freezing temperatures (−35°C) gave slightly higher degradability than higher temperatures. Untreated early season rice straw showed higher degradability than straw of middle and later season rice. There was a significant increase in the degradation of straw after urea treatment, and greatest for late and middle season rice straw. On average, urea-treatment of rice straw increased the DM and OM in sacco losses after 48 h of incubation (48 h) by 24.0% and 30.7%, respectively. In order to study the kinetics of the degradation of fibre fractions, the disappearance in sacco was also estimated for the loss of hemicellulose, cellulose and extractable biogenic silica (EBSi). There was a great variation in the content of silica between varieties. Rice straw degradation seemed to be related to the biogenic silica content (acid detergent insoluble silica (ADISi)). Urea treatment increased the extraction of biogenic silica and hence increased the degradation of hemicellulose and cellulose. The improvement in sacco disappearance of cellulose due to urea treatment was 36.8%, 19.5% and 5.3% for late, middle, and early rice straw, respectively. The degradability was higher for the stem than for the leaf blades and leaf sheaths. The response to urea treatment, however, was higher for leaf sheaths and leaf blades than for the stems, evening out differences in degradability. Urea treatment tended to increase the production of acetic acid whereas there was no effect on propionic and butyric acid production.     

Relative fibrolytic activities of anaerobic rumen fungi on untreated and sodium hydroxide treated barley straw in in vitro culture

The fibrolytic activities of rumen fungi were studied in terms of dry matter loss, plant cell wall degradation and enzyme (cellulase and xylanase) activities, when grown in vitro on either untreated or sodium hydroxide treated stems of barley straw over a 12 day period. Changes in fungal growth, development and overall biomass were followed using chitin assay and scanning electron microscopy. Treatment with sodium hydroxide resulted in a decrease in the NDF content together with the disruption of cuticle and the loosening and separation of the plant cells within the straw fragments. The enzyme activities of the anaerobic fungi have a high positive correlation (R(2)=0.99) with their biomass concentration assessed by chitin assay indicating that chitin is a valuable index for the estimation of the fungal biomass in vitro. The anaerobic fungi produced very extensive rhizoidal systems in these in vitro cultures. After incubation with rumen fungi, dry matter losses were, respectively, 35% and 38% for the untreated and treated straw samples and the overall fungal biomass, determined by chitin assay, was significantly higher in the treated samples. In vitro degradation of cellulose and hemicellulose was also higher in the treated than that of untreated cultures. Although, comparatively, xylanase activity was higher than that of cellulase, the cellulose fraction of the straw was degraded more than hemicellulose in both treated and untreated straw.     

Radiation and fermentation treatment of cellulosic wastes

The effect of radiation pasteurization of sugar cane bagasse and rice straw and fermentation using various strains of fungi were studied for upgrading of cellulosic wastes. The initial contamination by fungi and aerobic bacteria both in bagasse and straw was high. The doses of 30 kGy for sterilization and 8 kGy for elimination of fungi were required. Irradiation effect showed that rice straw contained comparatively radioresistant microorganisms. It was observed that all the fungi (Hericium erinacium, Pleurotus djamor, Ganoderma lucidum, Auricularia auricula, Lentinus sajor-caju, Coriolus versicolor, Polyporus arcularius, Coprinus cinereus) grow extending over the entire substrates during one month after inoculation in irradiated bagasse and rice straw with 3% rice bran and 65% moisture content incubated at 30°C. Initially, sugar cane bagasse and rice straw substrates contained 39.4% and 25.9% of cellulose, 22.9% and 26.9% of hemicellulose, and 19.6% and 13.9% of lignin + cutin, respectively. Neutral detergent fibre (NDF) values decreased significantly in sugar cane bagasse fermented byG. lucidum, A. auricula andP. arcularius, and in rice straw fermented by all the 8 strains of fungi. Acid detergent fibre (ADF) values also decreased in bagasse and rice straw fermented by all the fungi.P. arcularius, H. erinacium, G. lucidum andC. cinereus were found to be the most effective strains for delignification of sugar cane bagasse.     

Comparison of the rates of enzymatic hydrolysis of pretreated rice straw and bagasse with celluloses

The rates of enzymatic hydrolysis of pretreated rice straw and bagasse have been studied and compared with the hydrolysis rates of microcrystalline cellulose powder (MCCP) and Solka Floc. The effects of particle size reduction and enzyme loading on the rates of hydrolysis of rice straw and bagasse were also studied. It was found that the rates of hydrolysis of pretreated rice straw and bagasse are much higher than that of MCCP and Solka Floc. For both rice straw and bagasse, particle size reduction had very little effect in enhancing the rate of hydrolysis. Lignin present at <10% did not seem to hinder the accessibility of the enzyme to the cellulose surface. An enzyme loading > 40 Ug^?1 had no effect on the hydrolysis rate of rice straw or bagasse.     

不同木质纤维素基质上白腐菌降解特性的研究

通过测定木质素、纤维素、半纤维素和漆酶分泌的变化,研究白腐菌在稻草、木屑、粗纤维素、滤纸、黑液木素基质上的降解特性。结果表明,除黑液木素上白腐菌不能生长外,在前25d,各基质中纤维素、半纤维素和木质素含量呈持续下降趋势,之后,降解速率减少,其中木质素的降解速率大于纤维素和半纤维素的降解速率。漆酶分泌在生长初期呈快速上升趋势,第10d酶活达到最大,第10～20d快速下降,其后基本不变,基质中酶活大小顺序为稻草基质、木屑基质、粗纤维和滤纸基质,显示了木质素存在对漆酶分泌的诱导作用。     

Structure and saccharification of rice straw pretreated with sulfur trioxide micro-thermal explosion collaborative dilutes alkali

In this paper, a sulfur trioxide collaborative dilutes alkali method has been developed to pre-treat rice straw and it has been studied that the pre-treated rice straw structure affected the saccharification of the rice straw hydrolyzed by cellulose enzymatic hydrolysis. The results show that the reaction of the sulfur trioxide with rice straw resulted in the internal micro-thermal explosion, and the saccharification rate was 91% based on the pretreated rice straw with sulfur trioxide for 4h following 1% w/v NaOH treatment for 7h at 50°C.     

Characteristics of degraded cellulose obtained from steam-exploded wheat straw

The isolation of cellulose from wheat straw was studied using a two-stage process based on steam explosion pre-treatment followed by alkaline peroxide post-treatment. Straw was steamed at 200 degrees C, 15 bar for 10 and 33 min, and 220 degrees C, 22 bar for 3, 5 and 8 min with a solid to liquid ratio of 2:1 (w/w) and 220 degrees C, 22 bar for 5 min with a solid to liquid ratio of 10:1, respectively. The steamed straw was washed with hot water to yield a solution rich in hemicelluloses-derived mono- and oligosaccharides and gave 61.3%, 60.2%, 66.2%, 63.1%, 60.3% and 61.3% of the straw residue, respectively. The washed fibre was delignified and bleached by 2% H2O2 at 50 degrees C for 5 h under pH 11.5, which yielded 34.9%, 32.6%, 40.0%, 36.9%, 30.9% and 36.1% (% dry wheat straw) of the cellulose preparation, respectively. The optimum cellulose yield (40.0%) was obtained when the steam explosion pre-treatment was performed at 220 degrees C, 22 bar for 3 min with a solid to liquid ratio of 2:1, in which the cellulose fraction obtained had a viscosity average degree of polymerisation of 587 and contained 14.6% hemicelluloses and 1.2% klason lignin. The steam explosion pre-treatment led to a significant loss in hemicelluloses and alkaline peroxide post-treatment resulted in substantial dissolution of lignin and an increase in cellulose crystallinity. The six isolated cellulose samples were further characterised by FT-IR and 13C-CP/MAS NMR spectroscopy and thermal analysis.     

Continuous single cell protein production fromCellulomonas sp. andCandida utilis grown in mixture on barley straw

Summary Continuous fermentations with mixed cultures of the cellulolytic bacteriaCellulomonas sp. and the yeastCandida utilis were examined. Fermentations were carried out in an aerated 5-l fermenter with different preparations of wet disintegrated barley straw as the cellulose source (3.6–4.2%). The straw was pretreated with NaOH (3.2–8.5 kg NaOH/100 kg dry straw) under high pressure and temperature in a feedstuff pellet press. The quantity of dry cell mass produced and the breakdown of the straw were measured. Crude protein and ash content in cell dry matter and residual fiber were determined. The experiments showed thatCellulomonas sp. andCandida utilis may be grown together in a continuous culture (dilution rate D=0.12–0.14 h^–1) for at least 3 days without washing out one of the organisms. Highest productivity was 1.39 g cell dry matter/l/h when using straw pretreated with 5.7% NaOH. The dry cell product contained 58–66% crude protein and up to 51% of the organic fiber dry matter was solubilized. The yield constants were 0.32–0.61 g cell dry matter per g solubilized organic fibers.     

Biodegradation of the cellulose component of rice straw byPleurotus sajor-caju

Pleurotus sajor-caju is an efficient degrader of the cellulose component of rice straw. Protein contents of the biodegraded straw after fungal growth increased from 1.7 to 10.0 %. Crude fibre and cellulose contents decreased from 32.6 and 45.0 to 6.3 and 17.8 %, respectively. The in vivo digestibility of the biodegraded rice straw increased by 34.1 %.     

Mild alkaline pretreatment can achieve high hydrolytic and fermentation efficiencies for rice straw conversion to bioethanol

Abstract

Mild alkaline pretreatment was evaluated as a strategy for effective lignin removal and hydrolysis of rice straw. The pretreatment efficiency of different NaOH concentrations (0.5, 1.0, 1.5 or 2.0% w/w) was assessed. Rice straw (RS) pretreated with 1.5% NaOH achieved better sugar yield compared to other concentrations used. A cellulose conversion efficiency of 91% (45.84?mg/ml glucose release) was attained from 1.5% NaOH pretreated rice straw (PRS), whereas 1% NaOH pretreated rice straw yielded 35.10?mg/ml of glucose corresponding to a cellulose conversion efficiency of 73.81%. The ethanol production from 1% and 1.5% NaOH pretreated RS hydrolysates was similar at ～3.3% (w/v), corresponding to a fermentation efficiency of 86%. The non-detoxified hydrolysate was fermented using the novel yeast strain Saccharomyces cerevisiae RPP-03O without any additional supplementation of nutrients.     

Evaluation of effectiveness of pretreating rice straw with n-butylamine for improvement of sugar yield

Summary n-Butylamine (n-BA) pretreatment extracted a part of the holocellulose (cellulose and hemicellulose) in rice straw into the liquid phase, as a mixture of oligosaccharides and a small amount of monosaccharides. However, there was loss of sugars resulting mainly from decomposition of monomers by a long exposure to a high n-BA concentration. Approximately 70% of the total sugars in the pretreatment solutions was converted enzymatically into reducing sugars (mainly monomers). On the other hand, more than 90% of holocellulose in the residual rice straw after the optimal pretreatment was solubilized enzymatically in 120 h of reaction time with 10 w/v% substrate and 6 mg protein/ml cellulase. The main action of n-BA on rice straw was delignification, and highly crystalline cellulose was not swollen. Thus the enhancement of the enzymatic solubilization rate of rice straw appeared to be due to the increase of surface area accessible to the enzyme by delignification. It was demonstrated by the relationship of the vapour-liquid equilibrium of the n-BA/water system that the n-BA used was recovered easily. The loss of n-BA, at most, was 30% in the case of the weight ratio of n-BA to rice straw of 0.1.