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Recombinant beta-galactosidase derived from Enterobacter cloacae Zjut HJ2001 for efficient biotransformation of galactooligosaccharides 用于高效生物转化半乳寡糖的重组β-半乳糖苷酶,来源于泄殖腔肠杆菌 Zjut HJ2001
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-25 DOI: 10.1016/j.bej.2024.109514
Galactooligosaccharides (GOS), as a type of prebiotics, have excellent physical and chemical properties, and can be widely used in the pharmaceutical and food fields. Recently, the microbial β-galactosidases have gained widespread attentions in industrial GOS production. However, most β-galactosidases from microorganisms have low transgalactosylation activity, resulting in poor GOS yield of enzymatic transformation from lactose. In this paper, a brand new β-galactosidase derived from Enterobacter cloacae Zjut HJ2001 was screened out from soil, and successfully overexpressed, characterized, and mutated by combinatorial alanine-scanning and site-saturation mutagenesis. Compared to the yield of 51.73 % obtained by wild-type β-galactosidase with lactose concentration of 380 g/L, the obtained mutant β-gal-H542V achieved a higher GOS yield of 67.08 %, which was the highest in the reported literature. These results suggested that the developed mutagenesis strategy could improve the transgalactosylation efficiency, and the mutant β-gal-H542V could be regarded as a prospective biocatalyst for GOS industrial manufacturing.
半乳寡糖(GOS)作为一种益生元,具有优良的物理和化学特性,可广泛应用于医药和食品领域。近年来,微生物 β-半乳糖苷酶在工业化生产 GOS 方面受到广泛关注。然而,大多数微生物β-半乳糖苷酶的转半乳糖基化活性较低,导致从乳糖酶解转化的 GOS 产率较低。本文从土壤中筛选出了一种全新的β-半乳糖苷酶,并通过丙氨酸扫描和位点饱和突变的组合方法对其进行了成功的过表达、表征和突变。在乳糖浓度为 380 g/L 的条件下,野生型 β-半乳糖苷酶的产率为 51.73%,与之相比,所获得的突变体 β-gal-H542V 的 GOS 产率高达 67.08%,为文献报道的最高产率。这些结果表明,所开发的诱变策略可以提高转半乳糖基化的效率,突变体β-gal-H542V可被视为一种有望用于GOS工业生产的生物催化剂。
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引用次数: 0
Copper and zinc sulfides bioleaching by an extremely thermophilic archaeon in high NaCl concentration 高浓度氯化钠条件下极嗜热古生物对硫化铜和硫化锌的生物浸出作用
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-24 DOI: 10.1016/j.bej.2024.109509
Chloride bioleaching has received attention of several mineral processing industries, particularly in countries where there is scarcity of freshwater and only chloride-containing waters can be used. Therefore, the present work investigated the effect of NaCl (1.0 mol L−1) on the bioleaching of three sulfide minerals: chalcopyrite, bornite, and sphalerite by the thermophilic archaea Sulfolobus acidocaldarius. Chalcopyrite dissolution was only 25 % in the biotic experiment in the absence of chloride, but reached 90 % in the presence of both microorganisms and chloride, while less than 60 % extraction was observed in the abiotic experiment with chloride. In the experiments of bornite bioleaching, 86 % and 77 % of copper were extracted in the biotic and abiotic tests with chloride, respectively. In the absence of NaCl, the biotic and abiotic experiments presented similar copper dissolution (∼35 %). Finally, bioleaching experiments carried out with sphalerite showed zinc extractions below 35 % in all conditions tested. The main contribution from the archaea was its ability to produce low concentrations of ferric ion, which was partially precipitated as jarosite, resulting in low redox potential values (< 450 mV vs. Ag/AgCl), and efficiently bioleached bornite and chalcopyrite. Furthermore, XRD and SEM-EDS analyses demonstrated that sphalerite was practically not leached while bornite was transformed into new copper sulfide phases (CuS and Cu3FeS4). Jarosite and elemental sulfur were products of chalcopyrite and bornite bioleaching in the presence of chloride.
氯化物生物浸出已受到多个矿物加工行业的关注,尤其是在淡水匮乏且只能使用含氯化物水的国家。因此,本研究调查了 NaCl(1.0 mol L-1)对嗜热古菌 Sulfolobus acidocaldarius 对黄铜矿、辉锑矿和闪锌矿这三种硫化矿物进行生物浸出的影响。在无氯化物的生物实验中,黄铜矿的溶解度仅为 25%,但在微生物和氯化物同时存在的情况下,溶解度达到了 90%,而在有氯化物的非生物实验中,黄铜矿的提取率不到 60%。在波来石生物浸出实验中,在有氯化物的生物和非生物试验中,铜的萃取率分别为 86% 和 77%。在没有氯化钠的情况下,生物实验和非生物实验的铜溶解度相似(35%)。最后,用闪锌矿进行的生物浸出实验表明,在所有测试条件下,锌的提取率都低于 35%。古细菌的主要贡献在于其产生低浓度铁离子的能力,铁离子部分沉淀为绿泥石,导致低氧化还原电位值(< 450 mV vs. Ag/AgCl),并有效地生物浸蚀了辉铜矿和黄铜矿。此外,XRD 和 SEM-EDS 分析表明,闪锌矿几乎没有被浸出,而辉铜矿则转变成了新的硫化铜相(CuS 和 Cu3FeS4)。箭石和元素硫是黄铜矿和辉铜矿在氯化物存在下生物浸出的产物。
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引用次数: 0
Assessment of in situ product recovery techniques to enhance 2-phenylethanol production by Acinetobacter soli ANG344B 评估原位产品回收技术,以提高固氮醋酸杆菌 ANG344B 的 2-苯基乙醇产量
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-23 DOI: 10.1016/j.bej.2024.109508
The 2-phenylethanol (2-PE) production process by the newly isolated Acinetobacter soli ANG344B is limited by product toxicity. To overcome this limitation and enhance 2-PE production process, various alternatives based in in situ product removal (ISPR) approaches were evaluated. The approaches selected for assessment were gas stripping using the air supplied to the bioreactor, liquid-liquid extraction and adsorption. Adsorption was found to be the most promising approach to increase 2-PE production. Amberlite XAD 4 was chosen from the different adsorbents tested since it has high affinity for 2-PE, being able to adsorb 205.8 ± 8.1 mg2-PE/gdry resin. In a batch cultivation process, in presence of 3 % (dry w/v) of Amberlite XAD 4, A. soli ANG344B was able to produce 6.99 ± 0.06 g/L of 2-PE with a volumetric productivity of 0.17 ± 0.00 g/L.h, which represents an improvement of 3.3-fold. To the best of our knowledge, this is the highest 2-PE production reported for a wild-type bacteria. These findings highlight the potential of Acinetobacter soli ANG344B as 2-PE producer, contributing to the development of natural 2-PE production process.
新分离出的固态醋酸乙烯杆菌(Acinetobacter soli ANG344B)生产 2-苯基乙醇(2-PE)的过程受到了产物毒性的限制。为了克服这一局限性并改进 2-PE 生产工艺,我们评估了基于原位产物去除(ISPR)方法的各种替代品。选定用于评估的方法包括利用提供给生物反应器的空气进行气提、液液萃取和吸附。吸附法被认为是最有希望提高 2-PE 产量的方法。从测试的不同吸附剂中选择了 Amberlite XAD 4,因为它对 2-PE 有很高的亲和力,能够吸附 205.8 ± 8.1 mg2-PE/gdry 树脂。在批量培养过程中,在含有 3 %(干重/体积)Amberlite XAD 4 的情况下,A. soli ANG344B 能够生产 6.99 ± 0.06 g/L 的 2-PE,体积生产率为 0.17 ± 0.00 g/L.h,提高了 3.3 倍。据我们所知,这是野生型细菌的最高 2-PE 产量。这些发现凸显了单歧杆菌 ANG344B 作为 2-PE 生产者的潜力,有助于天然 2-PE 生产工艺的发展。
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引用次数: 0
Cell-free expression and biochemical characterization of polysaccharide-synthesizing glycosyltransferases 多糖合成糖基转移酶的无细胞表达和生化鉴定
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-23 DOI: 10.1016/j.bej.2024.109507
Polysaccharides like cellulose and hyaluronan are synthesized by membrane-bound family-2 glycosyltransferases (GTs) that play critical structural, metabolic, or functional roles in cells. Though GT-2 family has the maximum number of deposited gene sequences, the biochemistry is poorly understood due to enzyme production challenges. Here, we developed a cell-free expression (CFE) protocol to produce two GT-2 family representative cellulose synthase (PttCesA8 from Populus tremula x tremuloides) and hyaluronan synthase (SeHAS from Streptococcus equisimilis). The CFE products were obtained as reconstituted proteoliposomes directly at high yields and short processing times compared to other approaches. Enzyme expression was confirmed using SDS-PAGE and immunoblotting, while integration of GTs into lipid layers was observed using cryogenic electron microscopy. Both GTs were catalytically active with Michalis-Menten kinetic constants, Km for PttCesA8, was 295.8 µM, and SeHAS was 321.51 µM (toward UDP N-Acetyl Glucosamine) and 207.88 µM (toward UDP Glucuronic Acid), respectively, and with UDP inhibiting both GTs. Mutation of conserved residues in SeHAS also confirmed the importance of lysine-139, glutamine-248, and threonine-283 residues in hyaluronan biosynthesis. In summary, CFE methods enable expression of polysaccharide-synthesizing enzymes as proteoliposomes at high yields with relative ease for rapid biochemical and structural characterization studies.
纤维素和透明质酸等多糖是由膜结合的 2 族糖基转移酶(GTs)合成的,它们在细胞中发挥着关键的结构、代谢或功能作用。虽然GT-2家族拥有最多的基因序列,但由于酶的生产面临挑战,人们对其生物化学还知之甚少。在此,我们开发了一种无细胞表达(CFE)方案,用于生产两种 GT-2 家族的代表性纤维素合成酶(来自杨树 x tremuloides 的 PttCesA8)和透明质酸合成酶(来自马氏链球菌的 SeHAS)。与其他方法相比,CFE 产物以重组蛋白脂质体的形式直接获得,产量高,处理时间短。使用 SDS-PAGE 和免疫印迹法确认了酶的表达,同时使用低温电子显微镜观察了 GTs 与脂质层的整合。两种 GT 都具有催化活性,其 Michalis-Menten 动力常数为:PttCesA8 的 Km 为 295.8 µM,SeHAS 为 321.51 µM(对 UDP N-Acetyl 葡萄糖胺)和 207.88 µM(对 UDP 葡萄糖酸),UDP 对两种 GT 都有抑制作用。对 SeHAS 中保守残基的突变也证实了赖氨酸-139、谷氨酰胺-248 和苏氨酸-283 残基在透明质酸生物合成中的重要性。总之,CFE 方法能以蛋白脂质体的形式高产表达多糖合成酶,而且相对容易进行快速生化和结构表征研究。
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引用次数: 0
Extraction of poly(3-hydroxybutirate) from Priestia megaterium using non-halogenated solvents: A comparative performance analysis 使用非卤化溶剂从巨栉藻中提取聚(3-羟基丁酸酯):性能比较分析
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-21 DOI: 10.1016/j.bej.2024.109502
Solvent extraction using chloroform is the most common industrial process for poly(3-hydroxybutyrate) (P(3HB)) recovery from dried biomass, and still a major barrier to expanding the commercial application of this biodegradable biopolymer. Consequently, there is great interest in alternative non-halogenated solvents for this process and some relevant related results are available in the literature for P(3HB) recovery from Gram-negative bacteria. This work evaluated the potential of a set of non-halogenated solvents for the extraction of P(3HB) from Priestia megaterium, a Gram-positive bacterium of great potential for P(3HB) production. Ethyl acetate (EtAc), methyl ethyl ketone (MEK), dimethyl carbonate (DMC), dimethyl sulfoxide (DMSO), N,N-dimethylacetamide (NNDA), 2-heptanone (2-Hp), propylene carbonate (PC), and isoamyl propionate (IAP) were tested. Preliminary solubilization tests using commercial P(3HB) showed that EtAc, MEK, DMC and IAP had lower P(3HB) solubilization capacity (below 0.08 g/L for EtAc, MEK, and DMC; 1.3–2.5 g/L for IAP) than DMSO (65–70 g/L) and PC, 2-Hp and NNDA (>100 g/L). Then, only DMSO, PC, 2-Hp, and NNDA were evaluated in recovery tests with intracellular P(3HB). DMSO was not selective for P(3HB), causing digestion of cell wall components. PC, 2-Hp, and NNDA outperformed chloroform, but NNDA stood out for its remarkably higher recovery (98.5 %, 30 min, 140 ºC).
使用氯仿进行溶剂萃取是从干燥生物质中回收聚(3-羟基丁酸)(P(3HB))的最常见工业工艺,这仍然是扩大这种可生物降解的生物聚合物商业应用的主要障碍。因此,人们对这一工艺的非卤化溶剂替代品产生了浓厚的兴趣,文献中也有一些从革兰氏阴性细菌中回收 P(3HB) 的相关结果。这项工作评估了一组非卤代溶剂从巨头普氏菌(一种具有生产 P(3HB)巨大潜力的革兰氏阳性细菌)中提取 P(3HB)的潜力。对乙酸乙酯(EtAc)、丁酮(MEK)、碳酸二甲酯(DMC)、二甲基亚砜(DMSO)、N,N-二甲基乙酰胺(NNDA)、2-庚酮(2-Hp)、碳酸丙烯酯(PC)和丙酸异戊酯(IAP)进行了测试。使用商用 P(3HB)进行的初步增溶测试表明,EtAc、MEK、DMC 和 IAP 的 P(3HB)增溶能力(EtAc、MEK 和 DMC 低于 0.08 g/L;IAP 低于 1.3-2.5 g/L)低于 DMSO(65-70 g/L)和 PC、2-Hp 和 NNDA(100 g/L)。然后,仅对 DMSO、PC、2-Hp 和 NNDA 进行了细胞内 P(3HB) 恢复试验评估。DMSO 对 P(3HB) 没有选择性,会导致细胞壁成分被消化。PC、2-Hp 和 NNDA 的性能优于氯仿,但 NNDA 的回收率更高(98.5%,30 分钟,140 ºC)。
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引用次数: 0
Optimization of phenyllactic acid biosynthesis and separation by machine learning with neural network and overlay sampling uniform design 利用神经网络和叠加采样统一设计的机器学习优化苯乳酸的生物合成和分离
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-21 DOI: 10.1016/j.bej.2024.109506
Machine learning methodology with neural network models was developed using the datasets based on the overlay sampling uniform design (OSUD) for optimization of phenyllactic acid biosynthesis and separation processes by cryogels. Compared with the multiple regression, the machine learning models exhibited a significant improvement of predictive accuracy of phenyllactic acid biosynthesis, in which the radial basis function neural network (RBFNN) model had the best predictive performance with the accuracy increased by 65.2 %. The combination of RBFNN and OSUD was further employed to optimize the chromatographic separation of phenyllactic acid from crude fermentation broth using two poly(hydroxyethyl methacrylate) based anion-exchange cryogel packed-beds (grafted with (vinylbenzyl)trimethylammonium chloride and N,N-dimethylaminoethyl methacrylate). After optimizing the three critical separation parameters: sample volume (5.3–31.8 mL), flow velocity (1.0–6.0 cm/min), and elution salt concentration (0.05–0.3 mol/L), it was found that the models provided excellent predictions. The optimized recovery rates for the two packed-beds were determined to be 76.5 % and 83.0 %, and the optimal adsorption capacities were 0.26 mg/mL and 0.39 mg/mL from the fermentation broth, respectively. This study provides a reliable integrated approach for optimizing the synthesis and separation processes of high-value bioproducts like phenyllactic acid from crude feedstocks.
利用基于叠加抽样均匀设计(OSUD)的数据集开发了神经网络模型的机器学习方法,用于优化苯乳酸的生物合成和冷凝胶的分离过程。与多元回归相比,机器学习模型显著提高了苯乳酸生物合成的预测精度,其中径向基函数神经网络(RBFNN)模型的预测性能最佳,精度提高了 65.2%。研究人员进一步将 RBFNN 和 OSUD 结合使用,优化了使用两种聚(甲基丙烯酸羟乙酯)阴离子交换冷凝胶填料床(接枝有(乙烯基苄基)三甲基氯化铵和 N,N-二甲基氨基甲基丙烯酸甲酯)从粗发酵液中分离苯乳酸的色谱分离过程。在对样品体积(5.3-31.8 mL)、流速(1.0-6.0 cm/min)和洗脱盐浓度(0.05-0.3 mol/L)这三个关键分离参数进行优化后,发现模型提供了极好的预测。两种填料床的最佳回收率分别为 76.5 % 和 83.0 %,对发酵液的最佳吸附容量分别为 0.26 mg/mL 和 0.39 mg/mL。这项研究为优化从粗原料中合成和分离苯乳酸等高价值生物产品的过程提供了一种可靠的综合方法。
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引用次数: 0
Response of the performance and succession of denitrification consortium under the variation of nutritional conditions: Mechanisms and characteristics 营养条件变化对反硝化联合体性能和演替的影响:机理与特征
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-20 DOI: 10.1016/j.bej.2024.109503
In the process of wastewater treatment, biomass was typically subjected to variation of nutrient condition. This study investigated the effects of different nutrient condition on denitrification performance and bacterial communities. The results showed that under carbon-to-nitrogen ratio (C/N) was 4, denitrifying sludge (DS) exhibited excellent denitrification performance with influent nitrate (NO3--N) concentration in the range of 88.8 ± 9.04–297.5 ± 6.63 mg L−1, maintained total nitrogen removal efficiency (TNRE) of 98.1 ± 1.75 %. While, as influent NO3--N concentration reached 408.9 ± 9.56 mg L−1, the denitrification performance was inhibited, and this inhibition was reversible. Under appropriate substrate level (215.0 ± 13.01 mg L−1), the specific denitrification activity (SDA) elevated to 300 % of its original value. Additionally, under starvation stress, although the relative abundance of some starvation-resistant bacteria (such as Actinobacteriota and Lentimicrobium) increased, the TNRE decreased by 7.3 %. During the recovery phase, despite extracellular polymeric substances (EPS) promoted, the TNRE decreased to 78.3 ± 4.7 %. Starvation stress was less favorable to DS for activity recovery than that of feast condition. These findings contribute to elucidating the mechanisms for DS to respond of different nutrient condition.
在废水处理过程中,生物量通常会受到营养条件变化的影响。本研究探讨了不同营养条件对反硝化性能和细菌群落的影响。结果表明,在碳氮比(C/N)为 4 的条件下,反硝化污泥(DS)表现出优异的反硝化性能,进水硝酸盐(NO3--N)浓度范围为 88.8 ± 9.04-297.5 ± 6.63 mg L-1,总氮去除率(TNRE)保持在 98.1 ± 1.75 %。当进水 NO3-N 浓度达到 408.9 ± 9.56 mg L-1 时,反硝化性能受到抑制,而且这种抑制是可逆的。在适当的底物水平(215.0 ± 13.01 mg L-1)下,特定反硝化活性(SDA)提高到原来的 300%。此外,在饥饿胁迫下,虽然一些耐饥饿细菌(如放线菌和旬杆菌)的相对丰度有所增加,但 TNRE 却下降了 7.3%。在恢复阶段,尽管细胞外聚合物质(EPS)增加,TNRE 仍下降到 78.3 ± 4.7 %。与盛宴条件相比,饥饿胁迫不利于DS的活性恢复。这些发现有助于阐明 DS 对不同营养条件的响应机制。
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引用次数: 0
Economic validation and comparison of microbial tryptophan, erythritol and collagen production in an integrated sugarcane biorefinery 综合甘蔗生物精炼厂中微生物色氨酸、赤藓糖醇和胶原蛋白生产的经济验证与比较
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-19 DOI: 10.1016/j.bej.2024.109505

Low sugar prices present a significant challenge to the global sugarcane industry, prompting the exploration of diversification strategies for expanding product portfolios. Techno-economic analyses and environmental sustainability assessments were carried out to evaluate the microbial production of tryptophan, erythritol, and collagen from A-molasses in a biorefinery annexed to an existing sugarcane mill. Tryptophan production exhibited the highest profitability, with a minimum selling price (MSP) at 59.7 % of its current market price, although the achievable production volumes of tryptophan from one sugar mill would oversupply the global market. Due to the larger market size of for collagen the achievable production capacity in the collagen scenario would avoid market saturation, reducing the risk of oversupply and rendering it more economically viable. In contrast, erythritol production was marginally not profitable, with an MSP exceeding the current market price by 1 %, primarily attributed to high operational costs. All scenarios demonstrated relatively low greenhouse gas (GHG) emissions (ranging from 9.1 to 16.5 kg CO2eq/kg product), with tryptophan production emerging as the most environmentally favourable option due to minimal chemical and freshwater usage. When compared with literature-reported data on ethanol and short-chain fructooligosaccharides (scFOS), only collagen and ethanol production were deemed viable, based on their favourable profitability and contribution to the market.

低糖价格给全球甘蔗产业带来了巨大挑战,促使人们探索扩大产品组合的多样化战略。我们进行了技术经济分析和环境可持续性评估,以评估在现有甘蔗厂的附属生物精炼厂中用微生物从 A-淀粉中生产色氨酸、赤藓糖醇和胶原蛋白的情况。色氨酸生产的利润率最高,最低销售价格(MSP)为其当前市场价格的 59.7%,但一家糖厂可实现的色氨酸产量将使全球市场供过于求。由于胶原蛋白的市场规模较大,胶原蛋白方案中的可实现生产能力将避免市场饱和,降低供过于求的风险,使其更具经济可行性。相比之下,赤藓糖醇生产利润微薄,最大生产成本比当前市场价格高出 1%,主要原因是运营成本高。所有方案的温室气体(GHG)排放量都相对较低(从 9.1 到 16.5 千克 CO2eq/千克产品不等),色氨酸生产因化学品和淡水用量最少而成为最环保的方案。与乙醇和短链果寡糖(scFOS)的文献报告数据相比,只有胶原蛋白和乙醇生产因其有利可图和对市场的贡献而被认为是可行的。
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引用次数: 0
Evaluation on potential application of lipase immobilized on rice husks in enzymatic glycerolysis reaction 评估固定在稻壳上的脂肪酶在酶解甘油反应中的应用潜力
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-17 DOI: 10.1016/j.bej.2024.109500

Owing to high production cost and low reaction yield, immobilized lipase is rarely used in industrial glycerolysis. This research characterizes the performance of lipase immobilized on rice husk in glycerolysis reaction. By utilizing hexamethylenediamine (HMDA) and glutaraldehyde (GA) as coupling agents, lipase from Thermomyces lanuginosus was immobilized on oxidized rice husk (ORH). For comparison, another sample was prepared where the lipase was directly immobilized on ORH without the use of HMDA and GA. Then, monoglyceride production was performed via glycerolysis using the immobilized lipase. The FTIR analysis verify interactions on rice husk including rice husk oxidation, HMDA coupling, GA activation and lipase immobilization on rice husk. The study found that within the examined range of added lipase for immobilization (10–40 mg-protein/g-support), ORH–HMDA–GA–Lipase possessed superior outcomes in terms of protein loading, immobilization yield, and recovered glycerolysis activity compared to ORH–Lipase. Besides, ORH–HMDA–GA–Lipase exhibits better storage stability (60°C, 44.9 %) and higher reusability (90.0 % monoglyceride yield at the 8th cycle) against ORH–Lipase. The results confirm satisfying performance of the prepared immobilized lipase in glycerolysis and highlight its enhancements facilitated by coupling agents.

由于生产成本高、反应产率低,固定化脂肪酶很少用于工业甘油分解。本研究表征了固定在稻壳上的脂肪酶在甘油分解反应中的性能。通过使用六亚甲基二胺(HMDA)和戊二醛(GA)作为偶联剂,将来自嗜热酵母菌(Thermomyces lanuginosus)的脂肪酶固定在氧化稻壳(ORH)上。为了进行比较,还制备了另一种样品,将脂肪酶直接固定在 ORH 上,而不使用 HMDA 和 GA。然后,使用固定化脂肪酶通过甘油分解生产单甘酯。傅立叶变换红外分析验证了稻壳上的相互作用,包括稻壳氧化、HMDA偶联、GA活化和脂肪酶固定在稻壳上。研究发现,在所考察的固定化脂肪酶添加量范围内(10-40 毫克蛋白/克-支持物),ORH-HMDA-GA-脂肪酶在蛋白负载量、固定化产量和甘油分解活性恢复方面均优于 ORH-脂肪酶。此外,与 ORH-脂肪酶相比,ORH-HMDA-GA-脂肪酶具有更好的储存稳定性(60°C,44.9%)和更高的可重复使用性(第 8 个周期的单甘油酯产量为 90.0%)。这些结果证实了所制备的固定化脂肪酶在甘油分解方面具有令人满意的性能,并强调了偶联剂对其性能的促进作用。
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引用次数: 0
The solidification of heavy metal Pb2+-contaminated soil by enzyme-induced calcium carbonate precipitation combined with biochar 通过酶诱导碳酸钙沉淀结合生物炭固化重金属 Pb2+ 污染的土壤
IF 3.7 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-09-17 DOI: 10.1016/j.bej.2024.109496

The remediation of heavy metal Pb2+-contaminated soil by enzyme (urease)-induced calcium carbonate precipitation (EICP) combined with biochar was studied. The solidification efficiency of Pb2+ reached 98.41 % when the mass ratio of CaCl2/urea was 1:1 using EICP technology to remedy Pb2+-contaminated water. However, the formed precipitate was accompanied by unstable vaterite, and Pb2+ had the risk of secondary leaching. When the biochar of 5 wt% was added to the Pb2+-contaminated soil, the soil structure tended to be dense and the toxic leaching concentration of Pb2+ was less than 5 mg/L, which met the national standard of China. The addition of biochar increased the pH of the contaminated soil and changed the free Pb2+ into insoluble Pb(OH)2. The biochar provided more nucleation sites for urease, and part of Pb2+ were adsorbed on its surface or diffused into the pores of biochar, which effectively solidified Pb2+ in the soil.

研究了酶(脲酶)诱导碳酸钙沉淀(EICP)与生物炭相结合对重金属 Pb2+污染土壤的修复。当 CaCl2/ 尿素的质量比为 1:1 时,使用 EICP 技术处理 Pb2+ 污染的水,Pb2+ 的固化效率达到 98.41%。然而,形成的沉淀物伴随着不稳定的辉绿岩,Pb2+ 有二次沥滤的风险。在 Pb2+ 污染土壤中添加 5 wt%的生物炭后,土壤结构趋于致密,Pb2+ 的毒性浸出浓度小于 5 mg/L,达到了中国国家标准。生物炭的加入提高了污染土壤的 pH 值,使游离的 Pb2+ 转化为不溶性的 Pb(OH)2。生物炭为脲酶提供了更多的成核位点,部分 Pb2+ 被吸附在其表面或扩散到生物炭的孔隙中,从而有效固化了土壤中的 Pb2+。
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引用次数: 0
期刊
Biochemical Engineering Journal
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