提高木质纤维素酶解糖化效率的研究进展

木质纤维素高效水解为可发酵糖是其在生物燃料及高附加值化学品转化过程中至关重要的环节。从环保的角度出发,水解方法中的酶解是木质纤维素被彻底降解而无环境污染的有效途径,并且酶水解反应糖损耗低、副产物少、条件温和,因此受到广泛关注。但木质纤维素的组成与结构极为复杂,加之纤维素酶存在稳定性差、寿命短、活性低等缺陷,致使酶解效率较低,酶解糖化成本过高,为此国内外学者对如何提高木质纤维素酶解效率开展诸多方面的研究,综述了近年来提高木质纤维素酶解效率研究取得的最新理论研究及工艺进展,并就木质纤维素的预处理、产酶菌株/技术、酶复配/重组、酶解助剂、酶固定、外场作用、酶回收重利用及酶解反应器多方面的研究情况进行了总结,进一步展望了木质纤维酶解糖化的发展方向。     

解糖热解纤维素菌F32降解未预处理秸秆及产纤维素酶特性分析

【目的】明确极端嗜热厌氧木质纤维素降解菌解糖热解纤维素菌F32代谢特征,并分析其产酶特性。【方法】使用细胞计数法绘制菌株的生长曲线,使用离子色谱及气相色谱进行产物和残糖量分析,以DNS法及对硝基苯酚法检测菌株胞外蛋白的酶活性。【结果】解糖热解纤维素菌F32在以葡萄糖、微晶纤维素和未经预处理小麦秸秆为碳源时生长状况优于解糖热解纤维素菌DSM 8903。在以葡萄糖为碳源进行培养时,与菌株DSM 8903相比,菌株F32具有产乳酸较多,而产氢气较少的特点。在以微晶纤维素和未经预处理小麦秸秆为碳源进行培养时,与菌株DSM 8903相比,菌株F32胞外蛋白具有较高的内切纤维素酶活性和木聚糖酶活性。【结论】解糖热解纤维素菌F32表现出较强的木质纤维素降解能力,其与DSM 8903的产物组成及胞外蛋白的酶活性具有明显差异。     

表面活性剂对木霉菌产纤维素酶的影响

研究了TW80和“一枝花”洗衣粉作为表面活性剂对木霉菌纤维表酶发酵的影响。结果表明：当TW80在培养基中用量为0.2-0.5%时,对FPA最大提高率为29.6%,对木聚糖酶（Xylanase)最大提高率为31.1%,洗衣粉在培养基中用量为0.5%时,对促进木霉菌产Xylanase有较好的效果。最大提高率达34.4%。     

纤维素酶与木质纤维素生物降解转化的研究进展

利用纤维素酶将预处理后的秸秆降解成可发酵性单糖,然后发酵生产所需的液体燃料及化工产品的技术,对于我国解决能源、环境、人口就业等难题有着巨大的积极影响。在木质纤维素生物降解转化工艺中,减少纤维素酶用量及提高酶解效率是降低木质纤维素降解成本的关键。纤维素酶系和木质纤维素酶水解技术的改进需要深入了解纤维素酶系统的组成及其协同作用、纤维素酶的结构与功能以及纤维素酶的生产技术。将就以上几个方面的研究进展进行讨论,并深入探讨了纤维素酶糖化能力的评价方法。     

乙酸分级预处理甘蔗渣对纤维素酶解性能的影响

为提高甘蔗渣的纤维素酶解性能,采用乙酸脱木素结合碱脱乙酰基的预处理工艺 (Acetoline工艺) 对甘蔗渣进行预处理,考察了乙酸脱木素过程中若干因素对预处理结果的影响,并对预处理后甘蔗渣的纤维素酶解性能进行了研究。结果表明,经过Acetoline预处理后甘蔗渣在7.5％固体含量、15 FPU+10 CBU/g固体的纤维素酶和β-葡萄糖苷酶用量下酶解48 h,酶解聚糖转化率接近80％。与稀酸预处理相比,Acetoline预处理可以得到更高的酶解聚糖转化率。实验结果表明Acetoline工艺是一种可有效提高甘蔗渣纤维素酶解性能的预处理方法。     

稀酸和稀碱预处理对四种不同生物质资源制备还原糖的影响

本论文探讨了不同浓度的稀H_2SO_4和稀NaOH预处理对大豆秸秆、水稻秸秆、象草和狼尾草四种不同生物质酶解制备还原糖的影响。结果表明,大豆秸秆、水稻秸秆、象草和狼尾草具有较高的纤维素和半纤维素含量,是制备还原糖的理想原料。与稀H_2SO_4预处理相比,经稀NaOH预处理后的样品表现出较好的酶解性能。通过使用4%的NaOH对大豆秸秆和狼尾草进行预处理,还原糖产量分别为145.8 mg/mL和319.2 mg/mL。此外,以1%NaOH预处理后的水稻秸秆和象草为原料,可以分别获得385.2 mg/mL和231.6 mg/mL还原糖产量。     

白蚁消化系统转化和降解木质纤维素酶研究进展

木质纤维素是地球上最丰富的有机聚合物,白蚁是古老但进化最成功的高效木质纤维素降解者之一。了解白蚁降解高度抗性植物聚合物的机制对工业上生物质能源转化和生物仿生设计有重要的借鉴和指导价值。白蚁和其共生微生物产生的木质纤维素酶在其转化利用木质纤维素上发挥着重要作用。本文从来源作用方面对白蚁自身及其肠道原虫、细菌和真菌产生的纤维素酶、木聚糖酶和漆酶等酶研究概况进行了总结,对其存在的问题和前景进行了展望。本综述有助于全面了解白蚁消化系统木质纤维素酶的基因种类、来源、分布、表达以及酶活性和功能。     

木质纤维素原料酶水解产乙醇工艺的研究进展

木质纤维素原料预处理后,经水解、发酵等过程,可生产乙醇作为清洁燃料,这大大提高了农业和林业废弃物的利用率,减轻了环境污染,并为经济的可持续发展提供了保证。目前木质纤维素酶水解因其具有明显优势而受到重视,被普遍研究和采用。综述了近年来木质纤维素原料的预处理方法、酶与水解技术、发酵工艺以及发酵耦合分离技术的最新研究成果。     

木质纤维素酶基因资源挖掘及真菌酶系改造

简述了木质纤维素酶基因资源挖掘的策略和方法及其在丝状真菌酶系改造中的应用。从候选基因的获取(木质纤维素酶基因资源的挖掘和高效利用)、外源基因的表达、酶系的复配和重构等方面综述了丝状真菌酶系改造的最新进展,并提出了丝状真菌酶系改造中亟须解决的关键问题。     

生物表面活性剂修复重金属污染研究进展

重金属在环境中积累会对动植物和人体健康造成危害。生物表面活性剂环境相容性好,在环境污染修复方面的应用日益受到关注。本文介绍了生物表面活性剂及其在重金属污染修复中的应用;生物表面活性剂与重金属络合的机理;影响二者络合的因素(如pH值、表面活性剂浓度、重金属存在形态等);对生物表面活性剂修复重金属污染的前景进行了展望。     

The pattern of cell wall deterioration in lignocellulose fibers throughout enzymatic cellulose hydrolysis

Cell wall deterioration throughout enzymatic hydrolysis of cellulosic biomass is greatly affected by the chemical composition and the ultrastructure of the fiber cell wall. The resulting pattern of cell wall deterioration will reveal information on cellulose activity throughout enzymatic hydrolysis. This study investigates the progression and morphological changes in lignocellulose fibers throughout enzymatic hydrolysis, using (transmission electron microscopy) TEM and field emission scanning electron microscopy (FE‐SEM). Softwood thermo‐mechanical pulp (STMP) and softwood bleached kraft pulp (SBKP), lignocellulose substrates containing almost all the original fiber composition, and with lignin and some hemicellulose removed, respectively, was compared for morphology changes throughout hydrolysis. The difference of conversion between STMP and SBKP after 48 h of enzymatic hydrolysis is 11 and 88%, respectively. TEM images revealed an even fiber cell wall cross section density, with uneven middle lamella coverage in STMP fibers. SKBP fibers exhibited some spaces between cell wall and lamella layers due to the removal of lignin and some hemicellulose. After 1 h hydrolysis in SBKP fibers, there were more changes in the fiber cross‐sectional area than after 10 h hydrolysis in STMP fibers. Cell wall degradation was uneven, and originated in accessible cellulose throughout the fiber cell wall. FE‐SEM images illustrated more morphology changes in SBKP fibers than STMP fibers. Enzymatic action of STMP fiber resulted in a smoother fiber surface, along with fiber peeling and the formation of ribbon‐disjunction layers. SBKP fibers exhibited structural changes such as fiber erosion, fiber cutting, and fiber splitting throughout enzymatic hydrolysis. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012     

Novel Aspergillus hemicellulases enhance performance of commercial cellulases in lignocellulose hydrolysis

A novel hemicellulase-producing fungal strain was isolated from a local soil sample. The organism is identified as Aspergillus fumigatus based on ribosomal RNA analyses. The Aspergillus strain, designated as 2NB, produces both enzymes acting on xylan backbone (xylanase and β-xylosidase), and those acting on side chains (or accessory enzymes) notably α-arabinofuranosidase and acetyl-xylan esterase. The Asperigillus hemicellulases are characterized as having relatively low xylanase and β-xylosidase activities but high side chain removal activities. The activity ratio of side-chain acting enzymes to xylanase is higher than that of the Multifect enzyme, a commercial hemicellulase product. The potential of the novel hemicellulases in lignocelluloses bioprocessing was demonstrated with alkaline-pretreated switchgrass as lignocellulose substrate with hemicellulase supplemented with a ratio of xylanase activity to filter paper unit of 2:1. Supplement of Aspergillus hemicellulases to commercial cellulases significantly enhanced the hydrolysis of lignocellulose, achieving a 94% hydrolysis yield based on reducing sugar measurement, compared to 60% when no hemicellulase or 75% when Multifect enzyme was used under otherwise identical conditions. The significant improvement resulting from supplementing a hemicellulase mix with high side-chain removal activities suggests the importance of accessory hemicellulases in lignocellulose processing.     

Effect of surfactants on cellulose hydrolysis

The effect of surfactants on the heterogeneous enzymatic hydrolysis of Sigmacell 100 cellulose and of steam-exploded wood was studied. Certain biosurfactants (sophorolipid, rhamnolipid, bacitracin) and Tween 80 increased the rate of hydrolysis of Sigmacell 100, as measured by the amount of reducing sugar produced, by as much as seven times. The hydrolysis of steam-exploded wood was increased by 67% in the presence of sophorolipid. At the same time, sophorolipid was found to decrease the amount of enzyme adsorbed onto the cellulose at equilibrium. Sophorolipid had the greatest effect on cellulose hydrolysis when it was present from the beginning of the experiment and when the enzyme/cellulose ratio was low. (c) 1993 John Wiley & Sons, Inc.     

Ultrasound‐assisted dilute acid hydrolysis of tea processing waste for production of fermentable sugar

Lignocellulosic materials that are the most abundant plant biomass in the world have the potential to become sustainable sources of the produced value added products. Tea processing waste (TPW) is a good lignocellulosic source to produce the value added products from fermentable sugars (FSs). Therefore, the present study is undertaken to produce FSs by using ultrasound‐assisted dilute acid (UADA) and dilute acid (DA) hydrolysis of TPW followed by enzymatic hydrolysis. UADA hydrolysis of TPW was optimized by response surface methodology (RSM) at maximum power (900 W) for 2 h. The optimum conditions were determined as 50°C, 1:6 (w/v) solid:liquid ratio, and 1% (w/v) DA concentration, which yielded 20.34 g/L FS concentration. Furthermore, its DA hydrolysis was also optimized by using RSM for comparison and the optimized conditions were found as 120°C, 1:8 solid:liquid ratio, and 1% acid concentration, which produced 25.3 g/L FS yield. Even though the produced sugars with UADA hydrolysis are slightly less, but it can provide significant cost saving due to the lower temperature requirement and less liquid consumption. Besides, enzymatic hydrolysis applied after pretreatments of TPW were very more economic than the conventional enzymatic hydrolysis in the literature due to shorter time requiring. In conclusion, ultrasound‐assisted is a promising technology that can be successfully applied for hydrolysis of biomass and can be an alternative to the other hydrolysis procedures and also TPW can be considered as suitable carbon source for the production of value‐added products like biofuels, organic acids, and polysaccharides. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:393–403, 2016     

Effect of surfactants on separate hydrolysis fermentation and simultaneous saccharification fermentation of pretreated lodgepole pine

The effects of surfactants addition on enzymatic hydrolysis and subsequent fermentation of steam exploded lodgepole pine (SELP) and ethanol pretreated lodgepole pine (EPLP) were investigated in this study. Supplementing Tween 80 during cellulase hydrolysis of SELP resulted in a 32% increase in the cellulose‐to‐glucose yield. However, little improvement was obtained from hydrolyzing EPLP in the presence of the same amount of surfactant. The positive effect of surfactants on SELP hydrolysis led to an increase in final ethanol yield after the fermentation. It was found that the addition of surfactant led to a substantial increase in the amount of free enzymes in the 48 h hydrolysates derived from both substrates. The effect of surfactant addition on final ethanol yield of simultaneous saccharification and fermentation (SSF) was also investigated by using SELP in the presence of additional furfural and hydroxymethylfurfural (HMF). The results showed that the surfactants slightly increased the conversion rates of furfural and HMF during SSF process by Saccharomyces cerevisiae. The presence of furfural and HMF at the experimental concentrations did not affect the final ethanol concentration either. The strategy of applying surfactants in cellulase recycling to reduce enzyme cost is presented. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Optimization of enzyme complexes for lignocellulose hydrolysis

The ability of a commercial Trichoderma reesei cellulase preparation (Celluclast 1.5L), to hydrolyze the cellulose and xylan components of pretreated corn stover (PCS) was significantly improved by supplementation with three types of crude commercial enzyme preparations nominally enriched in xylanase, pectinase, and beta-glucosidase activity. Although the well-documented relief of product inhibition by beta-glucosidase contributed to the observed improvement in cellulase performance, significant benefits could also be attributed to enzymes components that hydrolyze non-cellulosic polysaccharides. It is suggested that so-called "accessory" enzymes such as xylanase and pectinase stimulate cellulose hydrolysis by removing non-cellulosic polysaccharides that coat cellulose fibers. A high-throughput microassay, in combination with response surface methodology, enabled production of an optimally supplemented enzyme mixture. This mixture allowed for a approximately twofold reduction in the total protein required to reach glucan to glucose and xylan to xylose hydrolysis targets (99% and 88% conversion, respectively), thereby validating this approach towards enzyme improvement and process cost reduction for lignocellulose hydrolysis.     

木质纤维素酸解副产物对D-乳酸生产菌Sporolactobacillus sp.Y2-8生长及发酵的影响

选择乙酸根、糠醛、5-羟甲基糠醛、苯酚、香草酸和丁香醛等6种典型木质纤维素酸解副产物,考察它们对D-乳酸生产菌Sporolactobacillus sp.Y2-8生长及发酵的影响。实验结果表明:酚类物质抑制作用最强烈,0.25 g/L丁香醛已经完全抑制了菌体的生长和D-乳酸的发酵;苯酚和香草酸在低浓度(≤1.0 g/L)时抑制作用较小,但质量浓度达到3 g/L时对D-乳酸产量的抑制率分别为99%和70%;3 g/L糠醛和5-羟甲基糠醛对产物的抑制率分别为60%与20%,抑制作用小于酚类;乙酸根的影响最小,10 g/L的乙酸钠对菌体的生长和发酵几乎无抑制作用;当抑制物混合时,存在着相互促进作用,抑制作用更强烈。     

Key technologies for bioethanol production from lignocellulose

Controversies on bioethanol produced from straw mainly revolve around the unfitted economical feasibility and environmental concerns of the process, which attribute mainly to unilateral researches from own specialties of each scholar without regard to the characteristics of the straws themselves. To achieve an economical and environmentally-friendly system of bioethanol production from straw, a number of breakthroughs are needed, not only in individual process steps, but also in the balance and combination of these processes. This article gives an overview of the new technologies required and the advances achieved in recent years, especial progresses achieved in our group, based on the concept of fractional conversions. An eco-industrial multi-production pattern is established, by which the maximum efficacy and benefit of process can be achieved due to the production of many high-value co-products simultaneously with ethanol. We believed that, in the future, the bioethanol production from straw will be competitive economically and environmentally.