Biologically Active Recombinant Proteins

A special issue of Bioengineering (ISSN 2306-5354). This special issue belongs to the section "Biochemical Engineering".

Deadline for manuscript submissions: closed (31 August 2023) | Viewed by 19594

Special Issue Editors

Institute of Biomedicine and Guangdong Provincial Key Laboratory of Bioengineering Medicine, Jinan University, Guangzhou 510630, China
Interests: therapeutic proteins; liposome-assisted drug delivery; bioactive peptides and protein; growth factors
Special Issues, Collections and Topics in MDPI journals
School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China
Interests: recombinant protein expression; protein expression systems; protein engineering; bacterial factory; bioactive materials
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Functional protein molecules have good biocompatibility, degradability and versatility, and have great application value in the fields of medicine, military and textiles. Oriented by clinical needs, the design of protein molecules, cell factory construction, protein adaptation and regulation and protein material processing applications can be improved to achieve targeted enhancement of protein functions and industrial production. Endowing proteins with intelligent controllability and performance upgrades in the production process is, therefore, a research hotspot.

This Special Issue on “Biologically Active Recombinant Proteins”, therefore, will focus on innovative, original research articles and comprehensive reviews that reflect the latest developments in the field. Both fundamental research and applied areas are welcome.

Submissions suitable for consideration may include, but are not limited to:

  1. Active peptides/protein biosynthesis research;
  2. Cell factories for active protein production;
  3. Protein purification and analysis;
  4. Protein-engineered functional materials;
  5. Construction and application of Bacterial-derived vectors for drug delivery;
  6. Protein based nanostructures for drug delivery;
  7. Peptide and protein nanotechnology.

Prof. Dr. Qi Xiang
Dr. Yi Ma
Guest Editors

Manuscript Submission Information

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Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • recombinant protein expression
  • cell factory
  • cell free synthesis
  • biomaterials
  • drug carrier
  • bacterial derivatives

Published Papers (10 papers)

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Research

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13 pages, 3372 KiB  
Article
A Thermally Stable Recombinant Human Fibronectin Peptide-Fused Protein (rhFN3C) for Faster Aphthous Ulcer (AU) Healing
by Xiang Cai, Jiawen Zhu, Xin Luo, Guoguo Jin, Yadong Huang and Lihua Li
Bioengineering 2024, 11(1), 38; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering11010038 - 29 Dec 2023
Viewed by 862
Abstract
Approximately 59.4–100% of head and neck cancer patients receiving radiotherapy or radio chemotherapy suffer from aphthous ulcers (AUs), which seriously affect the subsequent treatment. At the same time, AUs are a common oral mucosal disease with a high incidence rate among the population, [...] Read more.
Approximately 59.4–100% of head and neck cancer patients receiving radiotherapy or radio chemotherapy suffer from aphthous ulcers (AUs), which seriously affect the subsequent treatment. At the same time, AUs are a common oral mucosal disease with a high incidence rate among the population, often accompanied by severe pain, and affect both physical and mental health. Strategies to increase the ulcer healing rate and relieve pain symptoms quickly is a long-term clinical objective. Oral mucosal discontinuity is the main histological hallmark of AUs. So, covering the inner mucosal defect with an in vitro engineered oral mucosal equivalent shows good prospects for AU alleviation. Fibronectin (FN) is a glycopeptide in the extracellular matrix and exhibits opsonic properties, aiding the phagocytosis and clearance of foreign pathogens through all stages of ulcer healing. But native FN comes from animal blood, which has potential health risks. rhFN3C was designed with multi-domains of native FN, whose core functions are the recruitment of cells and growth factors to accelerate AU healing. rhFN3C is a peptide-fused recombinant protein. The peptides are derived from the positions of 1444–1545 (FNIII10) and 1632–1901 (FNIII12–14) in human native FN. We optimized the fermentation conditions of rhFN3C in E. coli BL21 to enable high expression levels. rhFN3C is thermally stable and nontoxic for L929, strongly promotes the migration and adhesion of HaCaT, decreases the incidence of wound infection, and shortens the mean healing time by about 2 days compared to others (p < 0.01). rhFN3C may have great potential for use in the treatment of AUs. The specific methods and mechanisms of rhFN3C are yet to be investigated. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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13 pages, 2567 KiB  
Article
Temperature-Controlled Expression of a Recombinant Human-like Collagen I Peptide in Escherichia coli
by Wenjie Xie, Qiqi Wu, Zhanpeng Kuang, Jianhang Cong, Qirong Zhang, Yadong Huang, Zhijian Su and Qi Xiang
Bioengineering 2023, 10(8), 926; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering10080926 - 04 Aug 2023
Cited by 2 | Viewed by 1370
Abstract
Collagen is the functional protein of the skin, tendons, ligaments, cartilage, bone, and connective tissue. Due to its extraordinary properties, collagen has a wide range of applications in biomedicine, tissue engineering, food, and cosmetics. In this study, we designed a functional fragment of [...] Read more.
Collagen is the functional protein of the skin, tendons, ligaments, cartilage, bone, and connective tissue. Due to its extraordinary properties, collagen has a wide range of applications in biomedicine, tissue engineering, food, and cosmetics. In this study, we designed a functional fragment of human type I collagen (rhLCOL-I) and expressed it in Escherichia coli (E. coli) BL21(DE3) PlysS containing a thermal-induced plasmid, pBV-rhLCOL-I. The results indicated that the optimal expression level of the rhLCOL-I reached 36.3% of the total protein at 42 °C, and expressed in soluble form. In a 7 L fermentation, the yield of purified rhLCOL-I was 1.88 g/L. Interestingly, the plasmid, pBV220-rhLCOL-I, was excellently stable during the fermentation process, even in the absence of antibiotics. Functional analyses indicated that rhLCOL-I had the capacity to promote skin cell migration and adhesion in vitro and in vivo. Taken together, we developed a high-level and low-cost approach to produce collagen fragments suitable for medical applications in E. coli. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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14 pages, 2690 KiB  
Article
Discovery and Characteristics of a Novel Antitumor Cyclopeptide Derived from Shark
by Fu Li, Minghua Lei, Junye Xie, Shujun Guo, Weicai Li, Xiujuan Ren, Teng Wang, Songxiong Lin, Qiuling Xie and Xiaojia Chen
Bioengineering 2023, 10(6), 674; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering10060674 - 01 Jun 2023
Viewed by 1122
Abstract
Peptides pose a challenge in drug development due to their short half-lives in vivo. In this study, we conducted in vitro degradation experiments on SAIF, which is a shark-derived peptide that we previously studied. The degradation fragments were sequenced and a truncated peptide [...] Read more.
Peptides pose a challenge in drug development due to their short half-lives in vivo. In this study, we conducted in vitro degradation experiments on SAIF, which is a shark-derived peptide that we previously studied. The degradation fragments were sequenced and a truncated peptide sequence was identified. The truncated peptide was then cloned and expressed via the E. coli system with traceless cloning to form a novel cyclic peptide in vitro oxidation condition via the formation of a disulfide bond between the N- and C-termini, which was named ctSAIF. ctSAIF exhibited high anti-HCC activity and enhanced enzymatic stability in vitro, and retained antitumor activity and good biocompatibility in systemic circulation in a HCC xenograft model. Our study discovered and characterized a novel shark-derived cyclic peptide with antitumor activity, laying a foundation for its further development as an antitumor drug candidate. The study also provided a new solution for peptide drug development. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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12 pages, 1885 KiB  
Article
A Novel Fusion Protein System for the Production of Nanobodies and the SARS-CoV-2 Spike RBD in a Bacterial System
by Dóra Nagy-Fazekas, Pál Stráner, Péter Ecsédi, Nóra Taricska, Adina Borbély, László Nyitray and András Perczel
Bioengineering 2023, 10(3), 389; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering10030389 - 22 Mar 2023
Cited by 2 | Viewed by 2112
Abstract
Antibodies are key proteins of the immune system, and they are widely used for both research and theragnostic applications. Among them, camelid immunoglobulins (IgG) differ from the canonical human IgG molecules, as their light chains are completely missing; thus, they have only variable [...] Read more.
Antibodies are key proteins of the immune system, and they are widely used for both research and theragnostic applications. Among them, camelid immunoglobulins (IgG) differ from the canonical human IgG molecules, as their light chains are completely missing; thus, they have only variable domains on their heavy chains (VHHs). A single VHH domain, often called a nanobody, has favorable structural, biophysical, and functional features compared to canonical antibodies. Therefore, robust and efficient production protocols relying on recombinant technologies are in high demand. Here, by utilizing ecotin, an Escherichia coli protein, as a fusion partner, we present a bacterial expression system that allows an easy, fast, and cost-effective way to prepare nanobodies. Ecotin was used here as a periplasmic translocator and a passive refolding chaperone, which allowed us to reach high-yield production of nanobodies. We also present a new, easily applicable prokaryotic expression and purification method of the receptor-binding domain (RBD) of the SARS-CoV-2 S protein for interaction assays. We demonstrate using ECD spectroscopy that the bacterially produced RBD is well-folded. The bacterially produced nanobody was shown to bind strongly to the recombinant RBD, with a Kd of 10 nM. The simple methods presented here could facilitate rapid interaction measurements in the event of the appearance of additional SARS-CoV-2 variants. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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13 pages, 2417 KiB  
Article
A Comparative Study of Morphology, Photosynthetic Physiology, and Proteome between Diploid and Tetraploid Watermelon (Citrullus lanatus L.)
by Zhanyuan Feng, Zhubai Bi, Dugong Fu, Lihan Feng, Dangxuan Min, Chensong Bi and He Huang
Bioengineering 2022, 9(12), 746; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9120746 - 01 Dec 2022
Cited by 1 | Viewed by 1594
Abstract
Watermelon is an important fruit that is widely distributed around the world. In particular, the production and consumption of watermelon in China ranks first in the world. Watermelon production is severely affected by a variety of biotic and abiotic stresses during cultivation, and [...] Read more.
Watermelon is an important fruit that is widely distributed around the world. In particular, the production and consumption of watermelon in China ranks first in the world. Watermelon production is severely affected by a variety of biotic and abiotic stresses during cultivation, and polyploidization can promote stress resistance and yield. However, the morphological and physiological characteristics of tetraploid watermelon and the underlying molecular mechanisms are still poorly understood. In this study, we revealed that the leaves, fruits, and seeds of tetraploid watermelon were significantly larger than those of the diploid genotype. Some physiological characteristics, including photosynthetic rate (Pn) and stomatal conductance (Gs), were greater, whereas the intercellular CO2 concentration (Ci) and transpiration rate (Tr) were lower in tetraploid than in diploid watermelon. Two-dimensional gel electrophoresis combined with tandem mass spectrometry (MALDI-TOF/TOF) was performed to compare proteomic changes between tetraploid and diploid watermelon. A total of 21 differentially expressed proteins were identified; excluding the identical proteins, 8 proteins remained. Among them, four proteins were upregulated and four were downregulated in tetraploid versus diploid genotypes. qRT-PCR results showed inconsistencies in gene expression and protein accumulation, indicating a low correlation between gene expression and protein abundance. Generally, this study extends our understanding of the traits and molecular mechanisms of tetraploid watermelon and provides a theoretical basis for watermelon polyploid breeding. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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12 pages, 2895 KiB  
Article
Recombinant Expression of Human IL-33 Protein and Its Effect on Skin Wound Healing in Diabetic Mice
by Yunxian Li, Shixin Lin, Sheng Xiong and Qiuling Xie
Bioengineering 2022, 9(12), 734; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9120734 - 28 Nov 2022
Cited by 5 | Viewed by 1815
Abstract
Chronic refractory wounds are one of the complications of diabetes mellitus that require effective therapy. The dermal-wound-healing property of IL-33 in diabetics is little understood. Therefore, this study aimed to express recombinant humanized mature IL-33 (rhmatIL-33) in Escherichia coli BL21 (DE3) and demonstrate [...] Read more.
Chronic refractory wounds are one of the complications of diabetes mellitus that require effective therapy. The dermal-wound-healing property of IL-33 in diabetics is little understood. Therefore, this study aimed to express recombinant humanized mature IL-33 (rhmatIL-33) in Escherichia coli BL21 (DE3) and demonstrate its efficacy on dermal wounds in streptozotocin (STZ)-induced diabetic and nondiabetic mice by the dorsal incisional skin wound model. Results revealed that the rhmatIL-33 accelerated the scratch-healing of keratinocytes and fibroblasts at the cellular level. The wounds of diabetic mice (DM) showed more severe ulceration and inflammation than wild-type mice (WT), and the exogenous administration of rhmatIL-33 increased wound healing in both diabetic and wild-type mice. Compared with the up-regulation of endogenous IL-33 mRNA after injury in WT mice, the IL-33 mRNA decreased after injury in DM mice. Exogenous IL-33 administration increased the endogenous IL-33 mRNA in the DM group but decreased the IL-33 mRNA expression level of the WT group, indicating that IL-33 plays a balancing role in wound healing. IL-33 administration also elevated ILC2 cells in the wounds of diabetic and non-diabetic mice and improve the transcript levels of YM1, a marker of M2 macrophages. In conclusion, Hyperglycemia in diabetic mice inhibited the expression of IL-33 in the dermal wound. Exogenous addition of recombinant IL-33 promoted wound healing in diabetic mice by effectively increasing the level of IL-33 in wound tissue, increasing ILC2 cells, and accelerating the transformation of macrophage M1 to M2 phenotype. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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21 pages, 5090 KiB  
Article
Efficient Robust Yield Method for Preparing Bacterial Ghosts by Escherichia coli Phage ID52 Lysis Protein E
by Yi Ma, Wenjun Zhu, Guanshu Zhu, Yue Xu, Shuyu Li, Rui Chen, Lidan Chen and Jufang Wang
Bioengineering 2022, 9(7), 300; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering9070300 - 07 Jul 2022
Cited by 4 | Viewed by 2236
Abstract
Bacterial ghosts (BGs) are nonliving empty bacterial shells without cytoplasm retaining original morphology and identical antigenicity of natural bacteria, making them high potential and promising vaccine candidates and delivery vehicles. However, the low yield of commonly used BGs preparation methods limits its mass [...] Read more.
Bacterial ghosts (BGs) are nonliving empty bacterial shells without cytoplasm retaining original morphology and identical antigenicity of natural bacteria, making them high potential and promising vaccine candidates and delivery vehicles. However, the low yield of commonly used BGs preparation methods limits its mass production and widely application. In order to improve BGs production, E. coli phage ID52 lysis protein E was introduced to generating BGs for the first time. Above all, we compared the lysis activity of lysis protein of E. coli phage φX174 and E. coli phage ID52 as well as the effects of promoters on the lysis activity of ID52-E, which shown that the lysis activity and BGs formation rate of protein ID52-E was significantly higher than protein φX174-E. Further, the lysis activity of ID52-E was significantly improved under the control of L-arabinose inducible promoter which initial induction OD600 reached as high as 2.0. The applicability of lysis protein ID52-E induced by L-arabinose was proved by preparing probiotic E. coli Nissle 1917 BGs and pathogenic Salmonella typhimurium BGs in mass production. This paper introduced a novel and highly efficient method for BGs preparation depending on recombinant expression of E. coli phage ID52-E under eco-friendly and reasonable price inducer L-arabinose. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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Review

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17 pages, 1869 KiB  
Review
Quenchbodies That Enable One-Pot Detection of Antigens: A Structural Perspective
by Hee-Jin Jeong
Bioengineering 2023, 10(11), 1262; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering10111262 - 30 Oct 2023
Cited by 1 | Viewed by 1062
Abstract
Quenchbody (Q-body) is a unique, reagentless, fluorescent antibody whose fluorescent intensity increases in an antigen-concentration-dependent manner. Q-body-based homogeneous immunoassay is superior to conventional immunoassays as it does not require multiple immobilization, reaction, and washing steps. In fact, simply mixing the Q-body and the [...] Read more.
Quenchbody (Q-body) is a unique, reagentless, fluorescent antibody whose fluorescent intensity increases in an antigen-concentration-dependent manner. Q-body-based homogeneous immunoassay is superior to conventional immunoassays as it does not require multiple immobilization, reaction, and washing steps. In fact, simply mixing the Q-body and the sample containing the antigen enables the detection of the target antigen. To date, various Q-bodies have been developed to detect biomarkers of interest, including haptens, peptides, proteins, and cells. This review sought to describe the principle of Q-body-based immunoassay and the use of Q-body for various immunoassays. In particular, the Q-bodies were classified from a structural perspective to provide useful information for designing Q-bodies with an appropriate objective. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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17 pages, 3482 KiB  
Review
β-Glucosidase and Its Application in Bioconversion of Ginsenosides in Panax ginseng
by Thi Ngoc Anh Tran, Jin-Sung Son, Muhammad Awais, Jae-Heung Ko, Deok Chun Yang and Seok-Kyu Jung
Bioengineering 2023, 10(4), 484; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering10040484 - 18 Apr 2023
Cited by 7 | Viewed by 1870
Abstract
Ginsenosides are a group of bioactive compounds isolated from Panax ginseng. Conventional major ginsenosides have a long history of use in traditional medicine for both illness prevention and therapy. Bioconversion processes have the potential to create new and valuable products in pharmaceutical [...] Read more.
Ginsenosides are a group of bioactive compounds isolated from Panax ginseng. Conventional major ginsenosides have a long history of use in traditional medicine for both illness prevention and therapy. Bioconversion processes have the potential to create new and valuable products in pharmaceutical and biological activities, making them both critical for research and highly economic to implement. This has led to an increase in the number of studies that use major ginsenosides as a precursor to generate minor ones using β-glucosidase. Minor ginsenosides may also have useful properties but are difficult to isolate from raw ginseng because of their scarcity. Bioconversion processes have the potential to create novel minor ginsenosides from the more abundant major ginsenoside precursors in a cost-effective manner. While numerous bioconversion techniques have been developed, an increasing number of studies have reported that β-glucosidase can effectively and specifically generate minor ginsenosides. This paper summarizes the probable bioconversion mechanisms of two protopanaxadiol (PPD) and protopanaxatriol (PPT) types. Other high-efficiency and high-value bioconversion processes using complete proteins isolated from bacterial biomass or recombinant enzymes are also discussed in this article. This paper also discusses the various conversion and analysis methods and their potential applications. Overall, this paper offers theoretical and technical foundations for future studies that will be both scientifically and economically significant. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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15 pages, 2032 KiB  
Review
Plant Exosome-like Nanoparticles as Biological Shuttles for Transdermal Drug Delivery
by Ye Wang, Yongsheng Wei, Hui Liao, Hongwei Fu, Xiaobin Yang, Qi Xiang and Shu Zhang
Bioengineering 2023, 10(1), 104; https://0-doi-org.brum.beds.ac.uk/10.3390/bioengineering10010104 - 12 Jan 2023
Cited by 9 | Viewed by 4063
Abstract
Exosomes act as emerging transdermal drug delivery vehicles with high deformability and excellent permeability, which can be used to deliver various small-molecule drugs and macromolecular drugs and increase the transdermal and dermal retention of drugs, improving the local efficacy and drug delivery compliance. [...] Read more.
Exosomes act as emerging transdermal drug delivery vehicles with high deformability and excellent permeability, which can be used to deliver various small-molecule drugs and macromolecular drugs and increase the transdermal and dermal retention of drugs, improving the local efficacy and drug delivery compliance. At present, there are many studies on the use of plant exosome-like nanoparticles (PELNVs) as drug carriers. In this review, the source, extraction, isolation, and chemical composition of plant exosomes are reviewed, and the research progress on PELNVs as drug delivery systems in transdermal drug delivery systems in recent years has elucidated the broad application prospect of PELNVs. Full article
(This article belongs to the Special Issue Biologically Active Recombinant Proteins)
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