Pichia pastoris production of a prolyl 4-hydroxylase derived from Chondrosia reniformis sponge: A new biotechnological tool for the recombinant production of marine collagen

Pozzolini, Marina; Scarfı̀, Sonia; Mussino, Francesca; Salis, Annalisa; Damonte, Gianluca; Benatti, Umberto; Giovine, Marco

doi:10.1016/j.jbiotec.2015.05.007

Cited by 17 publications

(10 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Then, to obtain a raw evaluation of the peptide sizes in the MCH solution mixture, an SDS gel electrophoresis was performed on the undigested, and trypsin-digested, collagen suspensions (Figure 1A). In particular, the electrophoretic analysis of a 7.5% polyacrylamide gel of the undigested collagen suspension (lane 3) revealed the presence of two bands of about 100 kDa corresponding to α1-fibrillar and α2-fibrillar collagen chains, and one band at 70 kDa that likely could correspond to the non-fibrillar collagen, which was identified, characterized, and obtained in recombinant form by some of us [21,22,23,48]. Conversely, in the trypsin-digested MCH solution mixture (lane 1 and lane 2, two different digestions), no protein bands were observable, indicating that in the established enzymatic conditions, a good degree of collagen digestion, and release of peptides with a molecular average size lower than 40 kDa, was obtained.…”

Section: Resultsmentioning

confidence: 99%

Elicited ROS Scavenging Activity, Photoprotective, and Wound-Healing Properties of Collagen-Derived Peptides from the Marine Sponge Chondrosia reniformis

et al. 2018

Self Cite

View full text Add to dashboard Cite

Recently, the bioactive properties of marine collagen and marine collagen hydrolysates have been demonstrated. Although there is some literature assessing the general chemical features and biocompatibility of collagen extracts from marine sponges, no data are available on the biological effects of sponge collagen hydrolysates for biomedical and/or cosmetic purposes. Here, we studied the in vitro toxicity, antioxidant, wound-healing, and photoprotective properties of four HPLC-purified fractions of trypsin-digested collagen extracts—marine collagen hydrolysates (MCHs)—from the marine sponge C. reniformis. The results showed that the four MCHs have no degree of toxicity on the cell lines analyzed; conversely, they were able to stimulate cell growth. They showed a significant antioxidant activity both in cell-free assays as well as in H2O2 or quartz-stimulated macrophages, going from 23% to 60% of reactive oxygen species (ROS) scavenging activity for the four MCHs. Finally, an in vitro wound-healing test was performed with fibroblasts and keratinocytes, and the survival of both cells was evaluated after UV radiation. In both experiments, MCHs showed significant results, increasing the proliferation speed and protecting from UV-induced cell death. Overall, these data open the way to the use of C. reniformis MCHs in drug and cosmetic formulations for damaged or photoaged skin repair.

show abstract

Section: Resultsmentioning

confidence: 99%

Elicited ROS Scavenging Activity, Photoprotective, and Wound-Healing Properties of Collagen-Derived Peptides from the Marine Sponge Chondrosia reniformis

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…P. pastoris has also been used for the production of hydroxylated nonfibrillar collagen from the marine sponge Chondrosia reniformis (Pozzolini et al, 2015). The genes encoding the α and ß subunits of P4H and collagen were all derived from this organism and were coexpressed in shake flask cultures of P. pastoris .…”

Section: Protein Polymers Produced In P Pastorismentioning

confidence: 99%

Production of protein-based polymers in Pichia pastoris

Werten

Eggink

Stuart

et al. 2019

Biotechnology Advances

View full text Add to dashboard Cite

Materials science and genetic engineering have joined forces over the last three decades in the development of so-called protein-based polymers. These are proteins, typically with repetitive amino acid sequences, that have such physical properties that they can be used as functional materials. Well-known natural examples are collagen, silk, and elastin, but also artificial sequences have been devised. These proteins can be produced in a suitable host via recombinant DNA technology, and it is this inherent control over monomer sequence and molecular size that renders this class of polymers of particular interest to the fields of nanomaterials and biomedical research. Traditionally, Escherichia coli has been the main workhorse for the production of these polymers, but the methylotrophic yeast Pichia pastoris is finding increased use in view of the often high yields and potential bioprocessing benefits. We here provide an overview of protein-based polymers produced in P. pastoris . We summarize their physicochemical properties, briefly note possible applications, and detail their biosynthesis. Some challenges that may be faced when using P. pastoris for polymer production are identified: (i) low yields and poor process control in shake flask cultures; i.e. , the need for bioreactors, (ii) proteolytic degradation, and (iii) self-assembly in vivo . Strategies to overcome these challenges are discussed, which we anticipate will be of interest also to readers involved in protein expression in P. pastoris in general.

show abstract

“…A newly discovered bacterial P4H found in Bacillus anthracis [56,57], might be able to overcome this limitation in future experiments. In addition, recent advances have been made in coexpressing eukaryotic proline-4-hydroxylase in origami-type E. coli [58], which provides conditions similar to that of the endoplasmic reticulum (ER) within its cytosol and therefore allow the activity of the transgenic P4H-complex, as well as the yeast P. pastoris [59,60] which, being an eukaryote, has the necessary organelles for early-stage collagen assembly and the ability to produce high MW proteins, although only with poor yields and varying degrees of hydroxylation [61,62].…”

Section: Collagen-analogues As Basis For Future Biomaterialsmentioning

confidence: 99%

Routes towards Novel Collagen-Like Biomaterials

Golser

Scheibel

2018

Fibers

View full text Add to dashboard Cite

Collagen plays a major role in providing mechanical support within the extracellular matrix and thus has long been used for various biomedical purposes. Exemplary, it is able to replace damaged tissues without causing adverse reactions in the receiving patient. Today's collagen grafts mostly are made of decellularized and otherwise processed animal tissue and therefore carry the risk of unwanted side effects and limited mechanical strength, which makes them unsuitable for some applications e.g., within tissue engineering. In order to improve collagen-based biomaterials, recent advances have been made to process soluble collagen through nature-inspired silk-like spinning processes and to overcome the difficulties in providing adequate amounts of source material by manufacturing collagen-like proteins through biotechnological methods and peptide synthesis. Since these methods also open up possibilities to incorporate additional functional domains into the collagen, we discuss one of the best-performing collagen-like type of proteins, which already have additional functional domains in the natural blueprint, the marine mussel byssus collagens, providing inspiration for novel biomaterials based on collagen-silk hybrid proteins.

show abstract

Pichia pastoris production of a prolyl 4-hydroxylase derived from Chondrosia reniformis sponge: A new biotechnological tool for the recombinant production of marine collagen

Cited by 17 publications

References 38 publications

Elicited ROS Scavenging Activity, Photoprotective, and Wound-Healing Properties of Collagen-Derived Peptides from the Marine Sponge Chondrosia reniformis

Elicited ROS Scavenging Activity, Photoprotective, and Wound-Healing Properties of Collagen-Derived Peptides from the Marine Sponge Chondrosia reniformis

Production of protein-based polymers in Pichia pastoris

Routes towards Novel Collagen-Like Biomaterials

Contact Info

Product

Resources

About