Reusable Floating Beads with Immobilized Xylose-Fermenting Yeast Cells for Simultaneous Saccharification and Fermentation of Lime-Pretreated Rice Straw

Novel bioreactor beads for simultaneous saccharification and fermentation (SSF) of lime-pretreated rice straw (RS) into ethanol were prepared. Genetically modified Saccharomyces cerevisiae cells expressing genes encoding xylose reductase, xylitol dehydrogenase, and xylulokinase were immobilized in calcium alginate beads containing inorganic lightweight filler particles to reduce specific gravity. For SSF experiments, the beads were floated in slurry composed of lime-pretreated RS and enzymes and incubated under CO₂ atmosphere to reduce the pH for saccharification and fermentation. Following this reaction, beads were readily picked up from the upper part of the slurry and were directly transferred to the next vessel with slurry. After 240 h of incubation, ethanol production by the beads was equivalent to that by free cells, a trend that was repeated in nine additional runs, with slightly improved ethanol yields. Slurry with pre-saccharified lime-pretreated RS was subjected to SSF with floating beads for 168 h. Although higher cell concentrations in beads resulted in more rapid initial ethanol production rates, with negligible diauxic behavior for glucose and xylose utilization, no improvement in the ethanol yield was observed. A fermentor-scale SSF experiment with floating beads was successfully performed twice, with repeated use of the beads, resulting in the production of 40.0 and 39.7 g/L ethanol. There was no decomposition of the beads during agitation at 60 rpm. Thus, this bioreactor enables reuse of yeast cells for efficient ethanol production by SSF of lignocellulosic feedstock, without the need for instruments for centrifugation or filtration of whole slurry.     

响应面法优化柠檬苦素降解酶的固定工艺

目的 采用响应面法优化柠檬苦素降解酶的固定工艺。方法 采用海藻酸钠-聚乙烯醇固定柠檬苦素降解酶，通过单因素和响应面实验对固定化条件进行优化，确定最佳的工艺参数。结果 优化后的工艺参数为：以聚乙烯醇(0.2 g/100 mL)-海藻酸钠(5.0 g/100 mL)为载体，戊二醛（2 mL/100 mL）为交联剂，采用包埋法固定柠檬苦素降解酶，其中粗酶液浓度0.06 mg/mL、CaCl2浓度6 mg/mL、固定化时间22 h。此条件下的柠檬苦素降解率为(97.87±0.32)%。结论 经优化后的工艺生产出的酶降解率高并且易于分离，具有很好的实际应用价值。     

京尼平交联制备枯草杆菌碱性蛋白酶聚集体

本研究利用新型交联剂京尼平制备了枯草杆菌碱性蛋白酶交联聚集体(BAP-CLEAs)。以酶活回收率为指标,确定了BAP-CLEAs制备的最佳条件为:交联剂质量浓度0.50%,交联温度35℃,交联时间12 h,此时BAP-CLEAs的酶活回收率为55.04%。采用扫描电镜及红外光谱对BAP-CLEAs进行表征,结果证明枯草杆菌碱性蛋白酶在京尼平的作用下成功交联。与游离酶相比,BAP-CLEAs的最适p H值向碱性方向偏移,由9.4变为10.3,在较宽的p H范围和温度范围内保持较高的酶活。另外,在2%浓度的酪蛋白底物中重复使用5次后,BAP-CLEAs还能保持86.42%的酶活性。以上催化特性的结果表明,枯草杆菌碱性蛋白酶在京尼平的作用下可成功交联形成酶聚集体,且该交联酶聚集体具有比游离酶更优越的p H稳定性、温度稳定性和重复使用稳定性,有良好的工业应用前景。     

醇脱氢酶在聚乙烯膜表面的固定化研究

以聚丙烯酸（PAA）改性的聚乙烯（PE）膜为载体，研究了醇脱氢酶（ADH）的两种固定化路线，并以甲醛为底物考察了固定化酶的催化性能。路线1用聚乙烯亚胺（PEI）进一步改性，使用戊二醛（GA）固定化ADH。最优固定化pH为6.0，温度为5～15℃，酶浓度为1.0 mg/ml，GA浓度为0.01%(质量)；固定化酶的最适反应pH为6.5，温度为15～30℃，反应速率最高为9.6 μmol/(L·min)；重复利用10次后可保持47.3%的活性。路线2以PAA-PE为载体，用1-(3-二甲氨基丙基)-2-乙基碳二亚胺盐酸盐（EDC）和N-羟基琥珀酰亚胺（NHS）为活化剂，固定化ADH。EDC和NHS最优摩尔比为1∶0.5，固定化时间为24 h；固定化酶的最适反应pH为6.5，温度为20～37℃，反应速率为15.58 μmol/(L·min)；重复利用10次后可保持53.8%的活性。     

In-situ immobilization of Sr radioactive isotope using nanocrystalline hydroxyapatite

Hydroxyapatite was used as the inert matrix for in-situ immobilization of strontium (Sr) radioactive isotopes at room temperature. A nano-emulsification method was applied to synthesize Sr-substituted calcium hydroxyapatite (Ca_1?xSr_x)₁₀(PO₄)₆(OH)₂. The concentration of incorporated Sr was in the range of 0 ≤ x ≤ 1. Immobilization of Sr was evaluated using a stable isotope instead of radioactive isotope. The effect of strontium concentration on the crystal structure was studied and the results have showed that in the whole concentration range, Sr forms solid solutions with the host hydroxyapatite crystal structure. Powders comprised of nanometre sized particles were obtained and their properties, such as crystallite and particle size, changes in lattice parameters as function of dopant content and thermal stability, were further examined. It was found that the crystal structure of obtained powders is thermally stable at high temperatures. No secondary phases were formed in as-prepared powders or during calcination. The results in this study showed that nano-emulsion strategy provides a simple pathway for synthesis of a single-phase Sr-substituted hydroxyapatite, which can be used for immobilization of Sr radioactive isotopes.