Cell-surface engineering of yeasts for whole-cell biocatalysts

Ye, Mengqi; Ye, Yuqi; Du, Zong‐Jun; Chen, Guanjun

doi:10.1007/s00449-020-02484-5

Cited by 28 publications

(16 citation statements)

References 159 publications

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“…However, on the other hand, enzymes located in the cell surface are similar to immobilized enzymes or cell surface-displaying enzymes, which not only were more stable than the same enzymes secreted in culture medium but also possessed excellent reusability via convenient centrifugation. 50,51 During the process of myrosinase reaction, temperature and pH are involved in the sulforaphene degradation and the rearrangement process, respectively. Accordingly, both reaction temperature and pH play vital roles in sulforaphene accumulation.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…These results further demonstrated that although the XPR2 signal peptide was involved in the secretion of the foreign enzyme, most of the recombinant myrosinase was retained by the yeast cells instead of being secreted into the supernatant. However, on the other hand, enzymes located in the cell surface are similar to immobilized enzymes or cell surface-displaying enzymes, which not only were more stable than the same enzymes secreted in culture medium but also possessed excellent reusability via convenient centrifugation. , …”

Section: Resultsmentioning

confidence: 99%

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Preparation of Sulforaphene from Radish Seed Extracts with Recombinant Food-Grade Yarrowia lipolytica Harboring High Myrosinase Activity

Wang

Jiang

Liang

et al. 2021

J. Agric. Food Chem.

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Sulforaphene prepared from glucoraphenin by myrosinase is one of the main active ingredients of radish, which has various biological activities and brilliant potential for food and pharmaceutical applications. In this paper, a recombinant food-grade yeast transformant 20-8 with high-level myrosinase activity was constructed by over-expressing a myrosinase gene from Arabidopsis thaliana in Yarrowia lipolytica. The highest myrosinase activity produced by the transformant 20-8 reached 44.84 U/g dry cell weight when it was cultivated in a 10 L fermentor within 108 h. Under the optimal reaction conditions, 6.1 mg of sulforaphene was yielded from 1 g of radish seeds under the catalysis of the crude myrosinase preparation (4.95 U) at room temperature within 1.5 h. What is more is that when the whole yeast cells harboring myrosinase activity were reused 10 times, the sulforaphene yield still reached 92.53% of the initial level. Therefore, this efficient approach has broad application prospects in recyclable and large-scale preparation of sulforaphene.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Preparation of Sulforaphene from Radish Seed Extracts with Recombinant Food-Grade Yarrowia lipolytica Harboring High Myrosinase Activity

Wang

Jiang

Liang

et al. 2021

J. Agric. Food Chem.

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“…Cell-surface display technology offers a novel and efficient technique to enhance whole-cell biocatalysts expressing heterologous proteins on the cell surface. 120 It permits the creation of targeting proteins or peptides on the microbic cell surface via fusing a suitable protein as an anchoring factor. Thereby, it combines protein purification, gene expression and lipase immobilization.…”

Section: Surface Display Techniquementioning

confidence: 99%

“…121 In this technique, the lipase is immobilized on the microbial cell wall automatically and genetically. 120 Lipase-displaying whole cells surfaces were found to be excellent biocatalysts owing to their lower cost of production, high efficiency and easy recycling process. 122 1n 1985, the technique of surface display was first discovered as phage display by Smith, when tiny proteins were displayed on the surface of the phage via fusion with capsid proteins.…”

Section: Surface Display Techniquementioning

confidence: 99%

“…Thereby, it combines protein purification, gene expression and lipase immobilization 121 . In this technique, the lipase is immobilized on the microbial cell wall automatically and genetically 120 . Lipase‐displaying whole cells surfaces were found to be excellent biocatalysts owing to their lower cost of production, high efficiency and easy recycling process 122 .…”

Section: Enzyme Immobilizationmentioning

confidence: 99%

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Recent developments of lipase immobilization technology and application of immobilized lipase mixtures for biodiesel production

Abdulmalek

Yan

2022

Biofuels Bioprod Bioref

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Lipases are considered the most widely used industrial biocatalysts, and play a significant role in diverse industrial and biotechnological applications such as in biodiesel. Biodiesel is a renewable and ecologically friendly biofuel, and has received significantly increased attention in recent years. The reaction media of biodiesel synthesis via the transesterification process of fats and oils could be considered cascade reactions. The use of a mixture of immobilized lipases as biocatalysts in biodiesel reactions has received much consideration since it is a ‘greener technique’ and has several advantages over free lipases. The selection of appropriate carrier materials and immobilization techniques fundamentally affects the immobilized lipases’ catalytic performance for biodiesel production. Hence, this review discusses the recent developments for carrier materials and techniques for lipase immobilization as well as the current status of immobilized lipases mixtures and their application for biodiesel synthesis. The initial part of this review presents a brief introduction of lipases and their application for biodiesel preparation. New carrier materials and different basic and recent lipase immobilization techniques are also evaluated. Most significantly, studies carried out on mixtures of immobilized lipase‐catalyzed biodiesel for the last 15 years (2006–2021) are summarized. The economic and environmental evaluation of enzymatic biodiesel production is also presented. In the future, the application of mixed and co‐immobilized lipases may be enlarged to other enzymes and processes. © 2022 Society of Chemical Industry and John Wiley & Sons, Ltd

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Measuring the capacity of yeast for surface display of cell wall‐anchored protein isoforms by using β‐lactamase as a reporter enzyme

Martinić Cezar,

Paić,

Prekpalaj

et al. 2024

FEBS Open Bio

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Yeast surface display is a promising biotechnological tool that uses genetically modified yeast cell wall proteins as anchors for enzymes of interest, thereby transforming yeast cell wall into a living catalytic material. Here, we present a comprehensive protocol for quantifying surface‐displayed β‐lactamase on the cell wall of model yeast Saccharomyces cerevisiae. We use β‐lactamase as a reporter enzyme, which we tagged to be anchored to the cell wall closer to its N or C terminus, through the portion of the Pir2 or Ccw12 cell wall proteins, respectively. The catalytic activity of surface‐displayed β‐lactamase is assessed by its ability to hydrolyze nitrocefin, which produces a colorimetric change that is quantitatively measured by spectrophotometric analysis at 482 nm. This system enables precise quantification of the potential of S. cerevisiae strains for surface display, continuous real‐time monitoring of enzyme activity, and facilitates the study of enzyme kinetics and interactions with inhibitors within the cell's native environment. In addition, the system provides a platform for high‐throughput screening of potential β‐lactamase inhibitors and can be adapted for the visualization of other enzymes, making it a versatile tool for drug discovery and bioprocess development.

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Cell-surface engineering of yeasts for whole-cell biocatalysts

Cited by 28 publications

References 159 publications

Preparation of Sulforaphene from Radish Seed Extracts with Recombinant Food-Grade Yarrowia lipolytica Harboring High Myrosinase Activity

Preparation of Sulforaphene from Radish Seed Extracts with Recombinant Food-Grade Yarrowia lipolytica Harboring High Myrosinase Activity

Recent developments of lipase immobilization technology and application of immobilized lipase mixtures for biodiesel production

Measuring the capacity of yeast for surface display of cell wall‐anchored protein isoforms by using β‐lactamase as a reporter enzyme

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