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The fed-batch production of mannosylerythritol lipids by Ustilago maydis DSM 4500 from hydrophilic carbon sources. Ustilago maydis DSM 4500 利用亲水性碳源批量生产甘露糖赤藓糖醇脂。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-09-21 DOI: 10.1007/s00449-024-03084-3
André D Valkenburg, George M Teke, Robert W M Pott, Eugéne van Rensburg

Glycolipids are a class of widely studied biosurfactants with excellent applicability in cosmetic and pharmaceutical formulations. This class of biosurfactants includes mannosylerythritol lipids (MELs), which have gained particular interest due to their moisturizing and healing activity for dry and damaged human skin, arising from conditions such as eczema. Traditionally, MELs have been produced by growing certain basidiomycetous yeasts on vegetable oils. However, oils are a comparatively expensive substrate, which negatively affects the economic performance of MEL production. In addition to this, vegetable oils significantly complicate the downstream processing required to produce a product with the required purity for most applications. To address these challenges, this study investigated MEL-A production exclusively from hydrophilic carbon sources by Ustilago maydis DSM 4500. By implementing a fed-batch production strategy, maximum MEL-A concentration of 0.87 g/L was achieved from glucose exclusively. Also, adding micronutrients (such as MnSO4) to MEL-A production showed a 24.1% increase in the product titer, implying other metabolites are formed, favoring MEL production.

糖脂是一类经过广泛研究的生物表面活性剂,在化妆品和药物配方中具有极佳的适用性。这类生物表面活性剂包括甘露糖基季戊四醇脂(MELs),由于其对干燥和受损的人体皮肤(如湿疹)具有保湿和修复活性,因此特别受到关注。传统上,MELs 是通过在植物油上培养某些基枝酵母而产生的。然而,植物油是一种相对昂贵的基质,这对 MEL 生产的经济效益产生了负面影响。此外,植物油还大大增加了生产大多数应用所需的纯度产品所需的下游加工的复杂性。为了应对这些挑战,本研究调查了 Ustilago maydis DSM 4500 完全利用亲水性碳源生产 MEL-A的情况。通过实施喂料批量生产策略,仅从葡萄糖中就获得了 0.87 克/升的最大 MEL-A 浓度。此外,在 MEL-A 生产过程中添加微量营养素(如 MnSO4)可使产品滴度提高 24.1%,这意味着其他代谢物的形成有利于 MEL 的生产。
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引用次数: 0
Encapsulation of Candida antarctica lipase B in metal-organic framework under ultrasound and using it to one-pot synthesis of 1,3,4,5-tetrasubstituted pyrazoles. 在超声波作用下在金属有机框架中封装白色念珠菌脂肪酶 B,并利用它一锅合成 1,3,4,5 四代吡唑。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-09-06 DOI: 10.1007/s00449-024-03083-4
Zeynab Rangraz, Ali Jafari, Mansour Shahedi, Mostafa M Amini, Zohreh Habibi

Encapsulating the enzyme in metal-organic frameworks (MOFs) is a convenient method to prepare MOF-enzyme biocomposite. In this study, Candida antarctica lipase B (CAL-B) was chosen to immobilize in Cu-BTC MOF under ultrasound irradiation. CAL-B was immobilized in Cu-BTC under ultrasound at 21 kHz and 11.4 W/cm2 and incubation. 98% of CAL-B was immobilized in Cu-BTC with 99 U/mg activity (threefold more active than the free CAL-B). The prepared biocomposite was characterized using FT-IR, XRD, TGA, SEM, EDX, and BET. The thermal and solvent stability of CAL-B@Cu-BTC was investigated. It was found that at a temperature of 55 ℃, CAL-B@Cu-BTC maintains its activity even after 2 h of incubation. Furthermore, in the presence of 20% and 50% concentrations of MeCN, THF, and DMF, CAL-B@Cu-BTC was found to have an activity of over 80%. A prepared biocatalyst was used to synthesize 1,3,4,5-tetrasubstituted pyrazole derivatives (50-75%) in a one-pot vessel, by adding phenyl hydrazine hydrochlorides, benzaldehydes, and dimethyl acetylenedicarboxylate.

将酶封装在金属有机框架(MOF)中是制备 MOF-酶生物复合材料的一种简便方法。本研究选择了白色念珠菌脂肪酶 B(CAL-B)在超声辐照下固定在 Cu-BTC MOF 中。在 21 kHz 和 11.4 W/cm2 的超声波和培养条件下,CAL-B 被固定在 Cu-BTC MOF 中。98% 的 CAL-B 被固定在 Cu-BTC 中,其活性为 99 U/mg (比游离的 CAL-B 活性高三倍)。利用傅立叶变换红外光谱(FT-IR)、X射线衍射(XRD)、热重分析(TGA)、扫描电镜(SEM)、电离辐射分析(EDX)和BET对制备的生物复合材料进行了表征。研究了 CAL-B@Cu-BTC 的热稳定性和溶剂稳定性。结果发现,在 55 ℃ 的温度下,CAL-B@Cu-BTC 即使在培养 2 小时后仍能保持其活性。此外,在 20% 和 50% 浓度的 MeCN、THF 和 DMF 存在下,CAL-B@Cu-BTC 的活性超过 80%。利用制备的生物催化剂,通过加入苯肼盐酸盐、苯甲醛和乙酰二甲酸二甲酯,在一锅容器中合成了 1,3,4,5-四取代的吡唑衍生物(50-75%)。
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引用次数: 0
A Review of Fucoxanthin Biomanufacturing from Phaeodactylum tricornutum. 从三 角藻中提取岩藻黄质生物制造技术综述。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-06-17 DOI: 10.1007/s00449-024-03039-8
Yunlong Pang, LiQin Duan, Bo Song, YuLin Cui, XiaoYong Liu, TingTing Wang

Microalgae, compared to macroalgae, exhibit advantages such as rapid growth rates, feasible large-scale cultivation, and high fucoxanthin content. Among these microalgae, Phaeodactylum tricornutum emerges as an optimal source for fucoxanthin production. This paper comprehensively reviews the research progress on fucoxanthin production using Phaeodactylum tricornutum from 2012 to 2022, offering detailed insights into various aspects, including strain selection, media optimization, nutritional requirements, lighting conditions, cell harvesting techniques, extraction solvents, extraction methodologies, as well as downstream separation and purification processes. Additionally, an economic analysis is performed to assess the costs of fucoxanthin production from Phaeodactylum tricornutum, with a comparative perspective to astaxanthin production from Haematococcus pluvialis. Lastly, this paper discusses the current challenges and future opportunities in this research field, serving as a valuable resource for researchers, producers, and industry managers seeking to further advance this domain.

与大型藻类相比,微藻类具有生长速度快、可进行大规模栽培、岩藻黄质含量高等优点。在这些微藻中,三尖杉藻(Phaeodactylum tricornutum)是生产狐黄素的最佳来源。本文全面回顾了2012年至2022年利用三 角藻生产岩藻黄质的研究进展,从菌株选择、培养基优化、营养要求、光照条件、细胞收获技术、提取溶剂、提取方法以及下游分离和纯化工艺等多个方面进行了详细阐述。此外,本文还进行了经济分析,以评估利用三 角藻生产岩藻黄质的成本,并与利用血球藻生产虾青素进行了比较。最后,本文讨论了这一研究领域当前面临的挑战和未来的机遇,为寻求进一步推动这一领域发展的研究人员、生产者和行业管理者提供了宝贵的资源。
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引用次数: 0
Isolation of native microorganisms from Shengli lignite and study on their ability to dissolve lignite. 从胜利褐煤中分离出原生微生物并研究其溶解褐煤的能力。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-08-10 DOI: 10.1007/s00449-024-03080-7
Yinglin Zhao, Ruina Chai, Jianguo Duan, Zhiying Yang, Shuheng Zhang, Yunying Liu, Xiaoxia Wang, Jianguo Cheng

To more greenly and efficiently utilize the abundant lignite resources and explore the microbial degradation and transformation potential of lignite for its environmentally friendly and resourceful utilization, Shengli lignite from the Hulunbuir region of Inner Mongolia, China, was selected as the research subject. Through the dilution plating method and streaking method, 31 native microorganisms were successfully isolated from the Shengli lignite, including 16 bacteria and 15 fungi. After microbial coal dissolution experiments, it was found that certain microorganisms have a significant dissolving effect on lignite, with some bacterial and fungal strains showing strong dissolution capabilities. In particular, the bacterium SH10 Lysinibacillus fusiformis and the fungus L1W Paecilomyces lilacinus demonstrated the best coal-dissolving abilities, with dissolution rates both reaching 60%. The products of microbial dissolution of lignite were analyzed using gas chromatography-mass spectrometry (GC-MS) technology, identifying a variety of small molecular organic compounds, including alkanes, alcohols, esters, and phenols. The results of this study provide a new perspective on the biodegradation of lignite and lay the foundation for the development of new lignite treatment and utilization technologies.

为了更加绿色、高效地利用丰富的褐煤资源,探索褐煤的微生物降解和转化潜力,实现褐煤的环境友好和资源化利用,研究人员选择了中国内蒙古呼伦贝尔地区的胜利褐煤作为研究对象。通过稀释平板法和条纹法,成功从胜利褐煤中分离出 31 种原生微生物,包括 16 种细菌和 15 种真菌。经过微生物溶煤实验发现,某些微生物对褐煤有明显的溶解作用,一些细菌和真菌菌株表现出很强的溶解能力。其中,细菌 SH10 Lysinibacillus fusiformis 和真菌 L1W Paecilomyces lilacinus 的溶煤能力最强,溶解率均达到 60%。利用气相色谱-质谱(GC-MS)技术分析了微生物溶解褐煤的产物,确定了多种小分子有机化合物,包括烷烃、醇、酯和酚。该研究结果为褐煤的生物降解提供了一个新的视角,为开发新的褐煤处理和利用技术奠定了基础。
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引用次数: 0
Developing endophytic Penicillium oxalicum as a source of lignocellulolytic enzymes for enhanced hydrolysis of biorefinery relevant pretreated rice straw. 开发草青霉内生菌作为木质纤维素分解酶的来源,以增强生物精炼相关预处理稻草的水解作用。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-09-09 DOI: 10.1007/s00449-024-03085-2
Gaurav Sharma, Baljit Kaur, Yashika Raheja, Amarjeet Kaur, Varinder Singh, Neha Basotra, Marcos Di Falco, Adrian Tsang, Bhupinder Singh Chadha

Endophytic fungi, as plant symbionts, produce an elaborate array of enzymes for efficient disintegration of lignocellulosic biomass into constituent monomeric sugars, making them novel source of lignocellulolytic CAZymes with immense potential in future biorefineries. The present study reports lignocellulolytic enzymes production potential of an endophytic halotolerant Penicillium oxalicum strain isolated from Citrus limon, under submerged and solid-state fermentation (SmF & SSF, respectively), in the presence and absence of salt (1 M NaCl). The comparative QTOF-LC/MS-based exoproteome analysis of the culture extracts unveiled differential expression of CAZymes, with the higher abundance of GH6 and GH7 family cellobiohydrolase in the presence of 1 M salt. The strain improvement program, employing cyclic mutagenesis and diploidization, was utilized to develop hyper-cellulase producing mutant strains of P. oxalicum. The enzyme production of the developed strain (POx-M35) was further enhanced through statistical optimization of the culture conditions utilizing glucose mix disaccharides (GMDs) as an inducer. This optimization process resulted in the lignocellulolytic cocktail that contained high titers (U/mL) of endoglucanase (EG) (146.16), cellobiohydrolase (CBHI) (6.99), β-glucosidase (β-G) (26.21), xylanase (336.05) and FPase (2.02 U/mL), which were 5.47-, 5.54-, 8.55-, 4.96-, and 4.39-fold higher when compared to the enzyme titers obtained in wild HP1, respectively. Furthermore, the lignocellulolytic cocktails designed by blending secretome produced by mutant POx-M35 with xylanases (GH10 and GH11) derived from Malbranchea cinnamomea resulted in efficient hydrolysis of unwashed acid pretreated (UWAP) rice straw slurry and mild alkali deacetylated (MAD) rice straw. This study underscores the potential of bioprospecting novel fungus and developing an improved strain for optimized production and constitution of lignocellulolytic cocktails that can be an important determinant in advancing biomass conversion technologies.

内生真菌是植物的共生体,能产生一系列复杂的酶,有效地将木质纤维素生物质分解成单体糖,是木质纤维素分解酶的新来源,在未来的生物炼制中具有巨大潜力。本研究报告了在有盐和无盐(1 M NaCl)条件下,通过浸没发酵和固态发酵(分别为 SmF 和 SSF),从柠檬中分离出的内生耐盐青霉菌株产生木质纤维素分解酶的潜力。基于 QTOF-LC/MS 的外蛋白组比较分析揭示了 CAZymes 的差异表达,在有 1 M 盐的情况下,GH6 和 GH7 家族的纤维生物水解酶含量更高。利用循环诱变和二倍体化的菌株改良计划,培育出了草甘膦高纤维素酶突变菌株。通过利用葡萄糖混合二糖(GMDs)作为诱导剂对培养条件进行统计优化,进一步提高了所培育菌株(POx-M35)的产酶量。这一优化过程产生的木质纤维素分解鸡尾酒含有高滴度(U/mL)的内切葡聚糖酶(EG)(146.16)、纤维生物水解酶(CBHI)(6.99)、β-葡萄糖苷酶(β-G)(26.21)、木聚糖酶(336.05)和 FPase(2.02 U/mL),分别比野生 HP1 的酶滴度高 5.47、5.54、8.55、4.96 和 4.39 倍。此外,通过将突变体 POx-M35 产生的分泌物与来自 Malbranchea cinnamomea 的木聚糖酶(GH10 和 GH11)混合而设计的木质纤维素分解鸡尾酒能高效水解未经酸洗预处理(UWAP)的水稻秸秆浆和轻度碱脱乙酰化(MAD)的水稻秸秆。这项研究强调了对新型真菌进行生物勘探和开发改良菌株的潜力,以优化木质纤维素分解鸡尾酒的生产和组成,这可能是推进生物质转化技术的一个重要决定因素。
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引用次数: 0
Evolution characteristics and molecular constraints of microbial communities during coal biogasification. 煤炭生物气化过程中微生物群落的演化特征和分子制约因素。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-09-27 DOI: 10.1007/s00449-024-03086-1
Tuo Shan, Yuan Bao, Xiangrong Liu, Xiaojing Wang, Dan Li

This study investigates the production of biomethane, and variation in microbial community and coal molecular structures using gas chromatography, 16S rRNA high-throughput sequencing and Fourier transform infrared spectroscopy. Additionally, the factors influencing microbial community structure at a molecular level are discussed. The results demonstrate that bituminous coal exhibits a higher biomethane yield than anthracite coal. In bituminous coal samples, Escherichia and Proteiniphilum are the predominant bacteria at day 0, while Macellibacteroides dominates from days 5 to 35. Methanofollis is the dominated archaea during days 0 to 15, followed by Methanosarcina on day 35. In anthracite coal samples, Soehngenia is the dominant bacterial genus at day 0; however, it transitions to mainly Soehngenia and Aminobacterium within days 5-15 before evolving into Acetomicrobium on day 35. Methanocorpusculum is predominantly found in archaeal communities during days 0-15 but shifts to Methanosarcina on day 35. Alpha diversity analysis reveals that bacterial communities have higher species abundance and diversity compared to archaeal communities. Redundancy analysis indicates a significant correlation between coal molecular structure and bacterial community composition (P value < 0.05), whereas no correlation exists with archaeal community composition (P value > 0.05). The research findings provide theoretical support for revealing the biological gasification mechanisms of coal.

本研究利用气相色谱法、16S rRNA 高通量测序法和傅立叶变换红外光谱法研究了生物甲烷的生产、微生物群落和煤分子结构的变化。此外,还讨论了在分子水平上影响微生物群落结构的因素。研究结果表明,烟煤的生物甲烷产量高于无烟煤。在烟煤样本中,第 0 天的主要细菌是埃希氏菌(Escherichia)和蛋白菌(Proteiniphilum),而第 5 天至第 35 天的主要细菌是麦氏菌(Macellibacteroides)。在第 0 至第 15 天,古细菌主要是甲壳动物,其次是第 35 天的甲壳动物。在无烟煤样本中,第 0 天的主要细菌属是 Soehngenia,但在第 5-15 天内主要转变为 Soehngenia 和 Aminobacterium,然后在第 35 天演变为 Acetomicrobium。在第 0-15 天期间,古细菌群落中主要是甲烷古细菌,但在第 35 天时则转变为甲烷弧菌。阿尔法多样性分析表明,与古菌群落相比,细菌群落具有更高的物种丰度和多样性。冗余分析表明,煤分子结构与细菌群落组成之间存在显著相关性(P 值为 0.05)。研究结果为揭示煤的生物气化机制提供了理论支持。
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引用次数: 0
Consolidated bioprocessing of lignocellulosic wastes in Northwest China for D-glucaric acid production by an artificial microbial consortium. 利用人工微生物联合体对中国西北地区木质纤维素废物进行综合生物处理,以生产 D-葡萄糖酸。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-08-19 DOI: 10.1007/s00449-024-03081-6
Hao Fang, Yuchen Li, Yuqi Song, Liang Yu, Xiangyang Song, Chen Zhao

D-glucaric acid is a platform chemical of great importance and the consolidated bioprocessing (CBP) of lignocellulose by the microbial consortium of Trichoderma reesei C10 and Saccharomyces cerevisiae LGA-1C3S2 features prospects in biomanufacturing it. Here we compared some representative lignocelluloses in Northwest China including corn stover, wheat straw and switchgrass, and the leading pretreatments including steam explosion, subcritical water pretreatment, sodium hydroxide pretreatment, aqueous ammonia pretreatment, lime pretreatment, and diluted sulfuric acid pretreatment. It was found that sodium hydroxide pretreated switchgrass (SHPSG) was the best substrate for D-glucaric acid production, resulting in the highest D-glucaric acid titers, 11.69 ± 0.73 g/L in shake flask and 15.71 ± 0.80 g/L in 10L airlift fermenter, respectively. To the best of our knowledge, this is the highest D-glucaric acid production titer from lignocellulosic biomass. This work offers a paradigm of producing low-cost D-glucaric acid for low-carbon polyethylene 2,5-furandicarboxylate (PEF) and a reference on developing biorefinery in Northwest China.

D-葡糖酸是一种非常重要的平台化学品,由毛霉C10和酿酒酵母LGA-1C3S2组成的微生物联合体对木质纤维素进行综合生物处理(CBP)具有生物制造木质纤维素的前景。在此,我们比较了西北地区具有代表性的木质纤维素(包括玉米秸秆、小麦秸秆和开关草)和主要预处理方法(包括蒸汽爆破、亚临界水预处理、氢氧化钠预处理、水氨预处理、石灰预处理和稀硫酸预处理)。研究发现,经氢氧化钠预处理的开关草(SHPSG)是生产 D-葡糖酸的最佳基质,其 D-葡糖酸滴度最高,在摇瓶中为 11.69 ± 0.73 g/L,在 10L 空运发酵罐中为 15.71 ± 0.80 g/L。据我们所知,这是利用木质纤维素生物质生产 D-葡糖酸的最高滴度。该研究为低碳聚 2,5-呋喃二甲酸乙二醇酯(PEF)生产低成本 D-葡糖酸提供了范例,也为中国西北地区发展生物炼制提供了参考。
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引用次数: 0
Enhancement of ε-poly-L-lysine production by Streptomyces albulus FQF-24 with feeding strategies using cassava starch as carbon source. 以木薯淀粉为碳源的喂养策略提高白链霉菌 FQF-24 的ε-聚-L-赖氨酸产量
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-12-01 Epub Date: 2024-08-16 DOI: 10.1007/s00449-024-03078-1
Boyan Li, Chenqi Wu, Senmeng Bai, Di Zhang, Chang Xu, Xiaofeng Yuan, Jiayi Tian, Jing Bai, Liangzhi Li, Jiaolong Fu

ε-Poly-L-lysine (ε-PL) is a natural and wide-spectrum antimicrobial additive. In this study, the production of ε-PL by Streptomyces albulus FQF-24 using cassava starch (CS) as carbon source and the effects of different feeding methods were investigated in a fermenter. The initial shake flask experiments demonstrated the efficient production of ε-PL with CS, achieving the ε-PL production of 1.18 g/L. Subsequent investigations in the fermenter identified that the ideal pH was 3.8 during the ε-PL synthesis phase. Under this condition, the production of ε-PL reached 1.35 g/L. When the pH was maintained at 3.8, the investigation of improvement of feeding composition was carried out in a 5 L fermenter. The intermittent feeding containing CS, inorganic and organic nitrogen sources resulted in the maximum ε-PL production and dry cell weight (DCW) reaching 17.17 g/L and 42.73 g/L. Additionally, continuous feeding with the composition of CS, organic and inorganic nitrogen sources, and inorganic salts further increased ε-PL production and DCW to 27.56 g/L and 38.5 g/L. Summarily, the above results indicate that the fermentation using low-cost CS and continuous feeding strategy with whole medium composition can provide a beneficial reference for the efficient production of ε-PL.

ε-聚-L-赖氨酸(ε-PL)是一种天然、广谱的抗菌添加剂。本研究以木薯淀粉(CS)为碳源,在发酵罐中研究了白链霉菌 FQF-24 生产ε-PL 的情况以及不同喂料方法的影响。最初的摇瓶实验表明,使用 CS 能高效生产ε-PL,ε-PL 产量达到 1.18 g/L。随后在发酵罐中进行的研究发现,ε-PL 合成阶段的理想 pH 值为 3.8。在此条件下,ε-PL 的产量达到了 1.35 克/升。当 pH 值保持在 3.8 时,在 5 升发酵罐中进行了改进投料成分的研究。含有 CS、无机氮和有机氮源的间歇喂料使ε-PL 产量和干细胞重量(DCW)达到最大值,分别为 17.17 克/升和 42.73 克/升。此外,连续饲喂含有 CS、有机和无机氮源以及无机盐成分的饲料可进一步提高ε-PL 产量和干细胞重量(DCW),分别达到 27.56 克/升和 38.5 克/升。总之,上述结果表明,采用低成本 CS 发酵和全培养基成分的连续饲喂策略可为高效生产ε-PL 提供有益的参考。
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引用次数: 0
Droplet-based bioprinting for the tailored fabrication of bacteria-laden living materials. 基于液滴的生物打印技术,用于定制制造含菌生物材料。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-22 DOI: 10.1007/s00449-024-03106-0
Xudong Guo, Dingyi Wang, Yingying Guo, Junpeng Zhang, Yingying Li, Haozhong Tian, Lihong Liu, Yong Liang, Yongguang Yin, Bin He, Ligang Hu, Guibin Jiang

Droplet-based bioprinting (DBB) allows for high precision, noncontact, and on-demand distribution of bioinks, hence it has been widely utilized in the preparation of bacteria-laden living materials (BLMs). Nonetheless, discontinuous ink deposition makes it challenging to fabricate large-sized intact living structures via this technique. Herein, we explore the way of using DBB to construct centimeter-scale BLMs with bespoke geometries, and further demonstrate its potential applicability in sensing-responsive device by integrating engineered bacteria. We first established a DBB method based on printing-path design, which does not require hardware modification. This strategy was able to produce customized 3D-hydrogel structures with high shape fidelity. Then, we confirmed the excellent biocompatibility of the above biofabrication approach. The Escherichia coli survived 93% ± 4.0% in printed BLMs, with uniform distribution throughout the structure. As a proof-of-concept, we finally manufactured a test strip-like heavy metal biosensor capable of plug-and-play detecting mercury (II) in water using the aforesaid approach. To our knowledge, this is the first study to employ 3D bioprinted BLMs for the detection of prevalent heavy metal pollutants. Our research shed light on the versatility of DBB in BLMs construction, which is not restricted to two-dimensional patterns. Moreover, our results are expected to innovate heavy metal biodetection and improve detection efficiency and sensitivity.

基于液滴的生物打印(DBB)可实现生物墨水的高精度、非接触和按需分布,因此已被广泛用于制备含菌生物材料(BLM)。然而,不连续的油墨沉积使得通过这种技术制备大尺寸的完整活体结构具有挑战性。在此,我们探索了使用 DBB 构建具有定制几何形状的厘米级 BLM 的方法,并通过整合工程细菌进一步证明了其在传感响应设备中的潜在适用性。我们首先建立了一种基于打印路径设计的 DBB 方法,这种方法不需要修改硬件。这种策略能够生产出形状保真度高的定制化三维水凝胶结构。然后,我们证实了上述生物制造方法具有良好的生物相容性。大肠杆菌在打印的 BLM 中存活率为 93% ± 4.0%,且在整个结构中分布均匀。作为概念验证,我们最终利用上述方法制造出了一种类似试纸的重金属生物传感器,能够即插即用地检测水中的汞(II)。据我们所知,这是第一项利用三维生物打印 BLM 检测普遍存在的重金属污染物的研究。我们的研究揭示了 DBB 在 BLM 构建中的多功能性,它并不局限于二维模式。此外,我们的研究成果有望创新重金属生物检测技术,提高检测效率和灵敏度。
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引用次数: 0
Enhancing D-pantothenate production in Escherichia coli through multiplex combinatorial strategies. 通过多重组合策略提高大肠杆菌的 D-泛酸产量。
IF 3.5 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2024-11-19 DOI: 10.1007/s00449-024-03105-1
Lianggang Huang, Landuo Sui, Yuan Yao, Yixuan Ma, Junping Zhou, Bo Zhang, Zhiqiang Liu, Yuguo Zheng

D-pantothenate, universally acknowledged as vitamin B5, has garnered considerable interest owing to its crucial functionality in the feed, pharmaceutical, and cosmeceutical sectors. Development of microbial strains for D-pantothenate hyperproducer has emerged as a prominent research direction in recent years. Herein, we converted an engineered Escherichia coli with low yield to a plasmid-free hyperproducer of D-pantothenate using multiplex combinatorial strategies. First, an initial strain was obtained through prolonging the cell lifespan. To promote the accumulation of D-pantothenic acid, the supply of cofactors was adaptively enhanced. Additionally, the heterologous gene panE from Pseudomonas aeruginosa, which encodes ketopantoate reductase (EC 1.1.1.169) catalyzing the synthesis of d-pantoate from α-ketopantoate, was screened and integrated into the chromosome. Subsequently, a strategy of acetate recycling and NOG pathway reconstruction were introduced and successfully to improve the D-pantothenate titer to 5.48 g/L. Additionally, we screened the regulatory factors and optimized its second codon to further increase the DPA yield of the engineered strains to 6.02 g/L in shake flask. The final engineered strain DS6 could efficiently produce 72.40 g/L D-pantothenate, which is 3.18-fold higher than the original strain. This study proposed a novel multiplex combination strategy for developing microbial cell factory of D-pantothenate, which was beneficial for the advancement of efficient D-pantothenate production.

D-泛酸(D-pantothenate)被普遍认为是维生素 B5,由于其在饲料、制药和化妆品领域的重要功能而备受关注。近年来,D-泛酸盐高产微生物菌株的开发已成为一个突出的研究方向。在此,我们采用多重组合策略,将产量低的工程大肠杆菌转化为无质粒的 D-泛酸盐高产菌株。首先,通过延长细胞寿命获得初始菌株。为了促进 D-泛酸的积累,适应性地加强了辅助因子的供应。此外,筛选出了铜绿假单胞菌的异源基因 panE,该基因编码酮泛酸还原酶(EC 1.1.1.169),可催化 α-酮泛酸合成 d-泛酸。随后,我们引入了醋酸盐循环和 NOG 途径重建策略,并成功地将 D-泛酸滴度提高到 5.48 g/L。此外,我们还筛选了调控因子并优化了其第二密码子,进一步将工程菌株在摇瓶中的 DPA 产量提高到 6.02 克/升。最终的工程菌株DS6可高效生产72.40 g/L的D-泛酸,是原始菌株的3.18倍。该研究提出了一种新型的多重组合策略,用于开发 D-泛酸盐微生物细胞工厂,有利于促进 D-泛酸盐的高效生产。
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Bioprocess and Biosystems Engineering
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