PBAT‒PLA膜袋受控需氧堆肥条件下的降解研究

目的 通过对广泛使用的PBAT–PLA生物降解膜袋在受控需氧堆肥条件下的降解机制研究，为生物降解塑料的大规模推广提供重要理论基础。方法 根据GB/T 19277.1—2011，在(58±2)℃需氧条件下，对PBAT–PLA膜袋进行为期160 d的生物降解测试(即工业堆肥)，并以常见的可降解材料微晶纤维素作为参比样品。对降解前后的材料进行红外、扫描电镜、能谱分析，并结合其所在堆肥样本的脂肪酶活性，从多角度探寻降解机制。结果 PBAT–PLA膜袋与微晶纤维素所在的堆肥脂肪酶活性都达到空白堆肥的3倍以上。红外显示由微晶纤维素水分子吸附、糖环打开、基团氧化形成的吸收峰加强，PBAT–PLA膜袋中的酯键峰明显减弱;扫描电镜发现降解的PBAT–PLA膜袋表面覆盖了微生物膜;能谱分析发现，碳元素大幅减少，氧元素增加。结论 微生物在PBAT–PLA膜袋表面生长形成生物膜，分泌大量脂肪酶，水解PBAT–PLA的酯键，使聚合物降解为不同链长的中间体或小分子，同时伴随着氧化，随后被作为碳源，在相关微生物体内被代谢利用，形成最终产物。     

Tempering–preservation treatment inactivated lipase in wheat bran and retained phenolic compounds

Wheat bran is rich in functional ingredients, but the high level of lipase limits its applications. Tempering–preservation treatment (at 70–90 °C with moisture of 20%–40% for 1–4 h) was exploited for stabilising wheat bran and its effect on polyphenols was investigated. The results showed that more lipase was inactivated at higher tempering moisture, temperature and longer time. The optimum condition for inactivation of wheat bran lipase was 30% moisture and 90 °C for 4 h. The inactivation rate reached 93.8% with a residual enzyme activity of 0.264 U g⁻¹. Under the optimum condition, the sum of free phenolic acids rose from 25.4 to 55.8 µg g⁻¹. As for bound phenolic acids, there was a slight increase of hydroxybenzoic acid derivatives but a slight decrease of hydroxycinnamic acid derivatives. The total contents of phenolic acids before and after stabilisation were not significantly different. This study showed the possibility of using tempering–preservation as an efficient method for inactivation of wheat bran lipase while maintaining its phenolic compounds, which could be used in the production of whole wheat flour.     

具脂肪酶和酯酶活性酵母菌的选育及酶学研究

从青岛市区含油土壤中筛选得到一株酵母 S9菌株 ,它可以同时产生较高酶活的脂肪酶和酯酶 .对影响 S9产脂肪酶和酯酶的各种因素进行了研究 ,1 %的可溶性淀粉为最佳碳源 ,最适的初始培养 p H和温度分别为 p H6.5和 2 8℃ ,对 S9脂肪酶和酯酶的各种酶学性质进行的研究表明 ,脂肪酶和酯酶的最适作用温度都为 3 0℃ ,最适作用 p H分别为 7.0和 8.0 ,两者都属于常温酶 ,在3 0～ 40℃范围内比较稳定 ,脂肪酶在中性偏碱的 p H环境中比较稳定 ,酯酶属于碱性酶 ,在 p H7.5～ 1 0 .5范围内相当稳定     

Kinetic study of liquid lipase-catalyzed glycerolysis of olive oil using Lipozyme TL 100L

Monoacylglycerol (MAG) and diacylglycerol (DAG) are two natural components found in most edible oils and fats. Conventional synthesis of MAG and DAG is usually conducted by glycerolysis of triacylglycerol (TAG) at high temperatures (above 200°C) in the presence of an alkaline catalyst. In this work, the synthesis of MAG and DAG using enzymatic glycerolysis of olive oil was investigated using Tween 80 as surfactant, n-butanol as co-surfactant and the novel lipase in free/liquid formulation Lipozyme TL 100L as catalyst. Experimental design was used to evaluate the effect of enzyme load and reaction temperature on the feedstock conversion. Enzyme load and system temperature were significant variables in the statistical design and the best condition was found at 35°C, 7.5 vol% of Lipozyme TL 100L and glycerol to oil volumetric ratio of 2:1 with conversion of TAG at approximately 98% after 2 h of process. A mathematical model based on the Ping-Pong Bi-Bi mechanism was used to describe the reaction kinetics. The model adequately described the behavior of the system and can be a useful tool for the design of reactors in larger scales.     

Typoselectivity of Crude Geobacillus sp. T1 Lipase Fused with a Cellulose-Binding Domain and Its Use in the Synthesis of Structured Lipids

Typoselectivity of crude CBD-T1 lipase (Geobacillus sp. T1 lipase fused with a cellulose binding domain) was investigated. Multi-competitive reaction mixtures including a set of n-chain fatty acids (C8:0, C10:0, C12:0, C14:0, C18:1 n-9, C18:2 n-6 and C18:3 n-3) and tripalmitin-enriched triacylglycerols were studied in hexane. The crude CBD-T1 lipase discriminated strongly against C18:1 n-9 [competitive factor (α) = 0.23] and showed the highest preference for C8:0 (α = 1). Utilizing the catalytic properties of crude CBD-T1 lipase, acidolysis of soybean oil with C8:0 was selected as a model reaction to investigate the ability of the lipase to produce MLM-type (medium-long-medium) structured lipids. Several reaction parameters (added water amount, reaction temperature, substrate molar ratio and reaction time) examined for incorporating C8:0 into soybean oil, the optimum conditions were: 1:3 (soybean oil/C8:0) of molar ratio, 3 mL of hexane, 50 °C of temperature, 48 h of reaction time, 20 % of crude CBD-T1 lipase (w/w total substrates), and 7.5 % of water (w/w enzyme). Under these conditions, the incorporation of C8:0 was 29.6 mol%. The results suggest that crude CBD-T1 lipase, which showed different fatty acid specificity profiles, is a potential biocatalyst for the modification of fats and oils.     

腾冲菌脂肪酶的表达纯化及酶学性质的表征

将来自腾冲菌的脂肪酶(Lip A)基因(lip A)克隆到大肠杆菌表达载体p ET28a(含T7启动子)和p Trc99A(含Trc启动子)中,转入大肠杆菌表达,发现Trc启动子更适合Lip A的表达。通过热处理和DEAE-Sepharose阴离子柱纯化过程,重组Lip A得到纯化,比酶活达到1.9U/mg,重组Lip A分子量为42k Da。重组Lip A在80℃、p H 4.5时酶活最高,经85℃保温2h,酶活保持60%以上;在p H值4.0~6.0之间,Lip A具有较好的稳定性;Cu2+和Zn2+对酶活力分别有38.9%和69.2%的抑制作用,Mn2+、Co2+和Tween-20对该酶有较大的激活作用;以p-nitrophenyl-laurate(p-NP-C12)为底物时,该酶的Km值为1.5mmol/L,kcat为34.5s-1。     

Enzymatic transesterification for production of biodiesel using yeast lipases: An overview

Biodiesel has provided an eco-friendly solution to fuel crisis, as it is renewable, biodegradable and a non-toxic fuel that can be easily produced through enzymatic transesterification of vegetable oils and animal fats. Enzymatic production of biodiesel has many advantages over the conventional methods as high yields can be obtained at low reaction temperatures with easy recovery of glycerol. Microbial lipases are powerful biocatalysts for industrial applications including biodiesel production at lower costs due to its potential in hydrolyzing waste industrial materials. Among them, lipases from yeasts, Candida antarctica, Candida rugosa, Cryptococcus sp., Trichosporon asahii and Yarrowia lipolytica are known to catalyze such reactions. Moreover, stepwise addition of methanol in a three step, two step and single step reactions have been developed using yeast lipases to minimize the inhibitory effects of methanol. The latest trend in biodiesel production is the use of whole-cell as biocatalysts, since the process requires no downstream processing of the enzyme. Synthesis of value added products from the byproduct glycerol further reduces the production cost of biodiesel. This review aims at compiling the information on various yeast lipase catalyzed transesterification reactions for greener production of biodiesel.     

Biochemical characterisation and application of lipases produced by Aspergillus sp. on solid‐state fermentation using three substrates

Lipase from Aspergillus sp. obtained by solid‐state fermentation (SSF) on wheat bran (LWB), soybean bran (LSB) and soybean bran combined with sugarcane bagasse (LSBBC) were 67.5, 58 and 57.3 U of crude lipase per gram substrate, respectively. The optimum pH of activity and stability of the LWB was between 8 and 9, and the optimum temperature of activity and stability was 50 °C and up to 60 °C, respectively. The LSB and LSBBC showed two peaks of optimum pH (4 and 6) and optimal values of temperature and stability at 50 °C. The LSB was stable in the pH range of 6–7, while LSBBC in the range of pH 4–7. All the enzymes show activities on p‐nitrophenyl esters (butyrate, laurate and palmitate). LWB stood out either on the hydrolysis of sunflower oil, presenting 66.1% of the activity over commercial lipase and on the esterification of oleic acid and ethanol, surpassing the activities of the commercial lipases studied. The thin layer chromatography showed that LWB and LSB have produced ethyl esters from corn oil, while LWB produced it from sunflower oil.