Recombinant Protein Production in Microalgae: Emerging Trends

2020 ◽  
Vol 27 (2) ◽  
pp. 105-110 ◽  
Author(s):  
Niaz Ahmad ◽  
Muhammad Aamer Mehmood ◽  
Sana Malik
Keyword(s):  
Chloroplast Genome ◽  
Recombinant Proteins ◽  
Large Scale ◽  
Higher Plants ◽  
Scale Up ◽  
Chloroplast Transformation ◽  
Point Of View ◽  
Nuclear Transformation ◽  
Scale Production ◽  
Algal Chloroplast

: In recent years, microalgae have emerged as an alternative platform for large-scale production of recombinant proteins for different commercial applications. As a production platform, it has several advantages, including rapid growth, easily scale up and ability to grow with or without the external carbon source. Genetic transformation of several species has been established. Of these, Chlamydomonas reinhardtii has become significantly attractive for its potential to express foreign proteins inexpensively. All its three genomes – nuclear, mitochondrial and chloroplastic – have been sequenced. As a result, a wealth of information about its genetic machinery, protein expression mechanism (transcription, translation and post-translational modifications) is available. Over the years, various molecular tools have been developed for the manipulation of all these genomes. Various studies show that the transformation of the chloroplast genome has several advantages over nuclear transformation from the biopharming point of view. According to a recent survey, over 100 recombinant proteins have been expressed in algal chloroplasts. However, the expression levels achieved in the algal chloroplast genome are generally lower compared to the chloroplasts of higher plants. Work is therefore needed to make the algal chloroplast transformation commercially competitive. In this review, we discuss some examples from the algal research, which could play their role in making algal chloroplast commercially successful.

Solid Lipid Nanoparticles: A Promising Drug Delivery Technology

2009 ◽  
Vol 2 (2) ◽  
pp. 509-516
Author(s):  
S. Pragati ◽  
S. Kuldeep ◽  
S. Ashok ◽  
M. Satheesh
Keyword(s):  
Drug Delivery ◽  
Solid Lipid Nanoparticles ◽  
Large Scale ◽  
Colloidal Particles ◽  
Scale Up ◽  
Lipid Nanoparticles ◽  
Scale Production ◽  
Research Activity ◽  
New Approach ◽  
Solid Lipid

One of the situations in the treatment of disease is the delivery of efficacious medication of appropriate concentration to the site of action in a controlled and continual manner. Nanoparticle represents an important particulate carrier system, developed accordingly. Nanoparticles are solid colloidal particles ranging in size from 1 to 1000 nm and composed of macromolecular material. Nanoparticles could be polymeric or lipidic (SLNs). Industry estimates suggest that approximately 40% of lipophilic drug candidates fail due to solubility and formulation stability issues, prompting significant research activity in advanced lipophile delivery technologies. Solid lipid nanoparticle technology represents a promising new approach to lipophile drug delivery. Solid lipid nanoparticles (SLNs) are important advancement in this area. The bioacceptable and biodegradable nature of SLNs makes them less toxic as compared to polymeric nanoparticles. Supplemented with small size which prolongs the circulation time in blood, feasible scale up for large scale production and absence of burst effect makes them interesting candidates for study. In this present review this new approach is discussed in terms of their preparation, advantages, characterization and special features.

Enhanced production of falcarinol-type polyacetylenes in hairy roots of cultivated carrot (Daucus carota subsp. sativus)

2021 ◽  
Author(s):  
Frank Dunemann ◽  
Christoph Böttcher
Keyword(s):  
Hairy Root ◽  
Large Scale ◽  
Higher Plants ◽  
Leaf Tissue ◽  
Hairy Root Cultures ◽  
Scale Production ◽  
Root Cultures ◽  
Tissue Samples ◽  
Positive Effects ◽  
Enhanced Production

Abstract Polyacetylenes (PAs) are a large group of bioactive phytochemicals, which are primarily produced by higher plants of the families Apiaceae and Araliaceae. Especially aliphatic C17-polyacetylenes of the falcarinol-type such as falcarinol (FaOH) and falcarindiol (FaDOH) are known for their numerous positive effects on human health. In this study we investigate the potential of carrot hairy root cultures for production of PAs. Three individual plants of seven differently coloured carrot cultivars were used for the development of hairy root cultures by transformation of root discs with the wild-type Rhizobium rhizogenes strain 15834. A total of 51 individual hairy root (HR) lines were obtained and quantitatively analysed together with root, petiole and leaf tissue samples for FaOH and FaDOH. Among the five tissues sampled from the donor plants, root periderm samples generally exhibited the highest PA levels with FaDOH as prevailing PA and large differences between cultivars. In comparison to periderm tissue, FaOH levels were highly increased in HR lines of all cultivars. In contrast, FaDOH levels were not significantly altered. Considering the low to moderate PA concentration in root and leaf tissues of the orange cultivars there was an up to more than 10-fold increase of the FaOH concentration in HRs of these genotypes. Within this study a reproducible method for Rrhi-mediated transformation of carrot root discs was applied which provides an efficient tool to assess the function of candidate genes involved in the biosynthesis of key PAs in carrot but might be used in future also for the large-scale production of falcarinol-type PAs.

SCREENING OF CHLOROPLAST PROMOTERS FOR HEVEA BRASILIENSIS CHLOROPLAST TRANSFORMATION

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Anas Akmal Ag. Ismail ◽  
Zaima Azira Zainal Abidin ◽  
Zarina Zainuddin
Keyword(s):  
Recombinant Protein ◽  
Plant Species ◽  
Recombinant Proteins ◽  
Hevea Brasiliensis ◽  
Agronomic Traits ◽  
Chloroplast Transformation ◽  
Foreign Protein ◽  
Protein Accumulation ◽  
Nuclear Transformation ◽  
Single Chain

In recent years, the growth in the use of recombinant proteins has grown tremendously. With the aid of the advances in DNA recombinant biotechnology, molecular farming in plants has been applied to meet this increasing demand where plants have emerged as one of the most promising general production platforms for recombinant proteins. Hevea brasiliensis is one of the main commodities in Malaysia and widely cultivated species for commercial production of latex. This important plant has been used to express recombinant proteins such as a single-chain variable fragment (scFv) antibody against the coat protein of Streptococcus gordonii (an oral dental bacterium), human serum albumin and human atrial natriuretic. The genes that encodes for the recombinant proteins were targeted into the nucleus genome of Hevea but the proteins were expressed in low concentration. Generating transgenic plant using chloroplast transformation offers many advantages in comparison to nuclear transformation and many researches have been made to apply this strategy to enhance agronomic traits or produce recombinant protein in several plant species. Since chloroplast is highly polyploidy, it allows high-level foreign protein expression. Given the generally very high foreign protein accumulation rates that can be achieved in transgenic chloroplasts, the aim of this study is to screen a number of chosen endogenous Hevea chloroplast promoters to drive the expression of the reporter gene, uidA for Hevea specific chloroplast transformation vector. Three promoters were chosen for this experiment which are; rbcL, psbA and rrn16 promoters. The putative regions of these promoters were derived from the chloroplast genome sequence of Hevea. Analyses of the three putative promoter regions using multiple sequence alignment with comparable regions from other plant species show significant sequence homology. Further analyses of the putative regions using in-vitro transcription are planned for future study. It is hoped that with the development of an optimized expression vector will allow high expression of valuable recombinant protein in the chloroplast of Hevea.

scholarly journals Scale-up of the production of highly reactive biogenic magnetite nanoparticles using Geobacter sulfurreducens

2015 ◽  
Vol 12 (107) ◽  
pp. 20150240 ◽  
Author(s):  
J. M. Byrne ◽  
H. Muhamadali ◽  
V. S. Coker ◽  
J. Cooper ◽  
J. R. Lloyd
Keyword(s):  
Magnetic Properties ◽  
Large Scale ◽  
Scale Up ◽  
Surface Reactivity ◽  
Geobacter Sulfurreducens ◽  
Scale Production ◽  
Buffer Solution ◽  
Pilot Plant Scale ◽  
Functional Biomaterials ◽  
Microbial Systems

Although there are numerous examples of large-scale commercial microbial synthesis routes for organic bioproducts, few studies have addressed the obvious potential for microbial systems to produce inorganic functional biomaterials at scale. Here we address this by focusing on the production of nanoscale biomagnetite particles by the Fe(III)-reducing bacterium Geobacter sulfurreducens , which was scaled up successfully from laboratory- to pilot plant-scale production, while maintaining the surface reactivity and magnetic properties which make this material well suited to commercial exploitation. At the largest scale tested, the bacterium was grown in a 50 l bioreactor, harvested and then inoculated into a buffer solution containing Fe(III)-oxyhydroxide and an electron donor and mediator, which promoted the formation of magnetite in under 24 h. This procedure was capable of producing up to 120 g of biomagnetite. The particle size distribution was maintained between 10 and 15 nm during scale-up of this second step from 10 ml to 10 l, with conserved magnetic properties and surface reactivity; the latter demonstrated by the reduction of Cr(VI). The process presented provides an environmentally benign route to magnetite production and serves as an alternative to harsher synthetic techniques, with the clear potential to be used to produce kilogram to tonne quantities.

scholarly journals GROWING OF VIRUS-FREE POTATO SEED TUBERS IN THE AEROPONIC PLANT

2020 ◽  
Vol 17 (35) ◽  
pp. 791-799
Author(s):  
Rafael ISMAGILOV ◽  
Ilgiz ASYLBAEV ◽  
Nuriya URAZBAKHTINA ◽  
Denis ANDRIYANOV ◽  
Firdavis AVSAKHOV
Keyword(s):  
Large Scale ◽  
Viral Infections ◽  
Potato Production ◽  
Poor Quality ◽  
Point Of View ◽  
Potato Seed ◽  
Scale Production ◽  
Planting Material ◽  
Large Scale Production

Throughout the world, potatoes, as a food crop, are very important. One of the main reasons for the poor quality of planting material, yield and potatoes themselves are viral infections. The use of virus-free seed material is one of the high-potential ways to increase the yield and efficiency of potato production. Aeroponics is a promising direction in obtaining a virus-protected crop. This study aimed to assess the potential and improve the technology for growing healthy mini-tubers of potatoes using the aeroponic method, which is a safe and economical method. Compared to the usual method of growing crops, aeroponics assumes lower water and energy costs per unit of production, as well as excludes soil diseases of the plant and does not allow damage to the tuber caused by pests. For growing different varieties of crops in different regions, artificial conditions such as additional lighting in greenhouses can be easily provided. In this study, economic calculations have shown that, from a practical point of view, Aeroponics technology may be appropriate for large-scale production of seed potatoes.

Identification of Polycistronic Transcriptional Units and Non-canonical Introns in Green Algal Chloroplasts Based on Long-read RNA Sequencing Data

2020 ◽  
Author(s):  
Xiaoxiao Zou ◽  
Heroen Verbruggen ◽  
Tianjingwei Li ◽  
Jun Zhu ◽  
Zuo Chen ◽  
...  
Keyword(s):  
Chloroplast Genome ◽  
Green Algae ◽  
Higher Plants ◽  
Green Algal ◽  
Chloroplast Genomes ◽  
Caulerpa Lentillifera ◽  
Transcriptional Units ◽  
Group Ii ◽  
Distinct Features ◽  
Algal Chloroplast

Abstract Background: Chloroplasts are important semi-autonomous organelles in plants and algae. Unlike higher plants, the chloroplast genomes of green algal linage have distinct features both in organization and expression. Despite the architecture of chloroplast genome have been extensively studied in higher plants and several model species of algae, little is known about transcriptional features in green algal lineages. Results: Based on full-length cDNA (Iso-Seq) sequencing, we identified widely co-transcribed polycistronic transcriptional units (PTUs) in the green alga Caulerpa lentillifera. In addition to clusters of genes from the same pathway, we identified a series of PTUs of up to nine genes whose function in the plastid is not understood. The RNA data further allowed us to confirm widespread expression of fragmented genes and conserved open reading frames, which are both important features in green algal chloroplast genomes. In addition, a newly fragmented gene specific to C. lentillifera was discovered, which may represent a recent gene fragmentation event in chloroplast genome.Taking the accurate exon-intron boundary information, gene structural annotation was greatly improved across the siphonous green algae lineages. Our data also revealed a type of non-canonical Group II introns, with a deviant secondary structure and intronic ORFs lacking known splicing or mobility domains. These widespread introns have conserved positions in their genes and are excised precisely despite lacking clear consensus intron boundaries.Conclusion: Our study fills important knowledge gaps in chloroplast genome organization and transcription in green algae, and providing new insights into expression of polycistronic transcripts, freestanding ORFs and fragmented genes in algal chloroplast genomes. Moreover, we revealed an unusual type of Group II intron with distinct features and conserved positions in Bryopsidales. Our data represents interesting additions to knowledge of chloroplast intron structure and highlights clusters of uncharacterized genes that probably play important roles in plastid.

Plants as factories for production of biopharmaceutical and bioindustrial proteins: lessons from cell biologyThis review is one of a selection of papers published in the Special Issue on Plant Cell Biology.

2006 ◽  
Vol 84 (4) ◽  
pp. 679-694 ◽  
Author(s):  
Allison R. Kermode
Keyword(s):  
Recombinant Proteins ◽  
Protein Function ◽  
Cell Biology ◽  
Large Scale ◽  
Production Systems ◽  
Plant Cells ◽  
Scale Production ◽  
Functional Protein ◽  
Large Scale Production ◽  
Engineer Plant

Transgenic plants, seeds, and cultured plant cells are potentially one of the most economical systems for large-scale production of recombinant proteins for industrial and pharmaceutical uses. Biochemical, technical, and economic concerns with current production systems have generated enormous interest in developing plants as alternative production systems. However, various challenges must be met before plant systems can fully emerge as suitable, viable alternatives to current animal-based systems for large-scale production of biopharmaceuticals and other products. Aside from regulatory issues and developing efficient methods for downstream processing of recombinant proteins, there are at least two areas of challenge: (1) Can we engineer plant cells to accumulate recombinant proteins to sufficient levels? (2) Can we engineer plant cells to post-translationally modify recombinant proteins so that they are structurally and functionally similar to the native proteins? Attempts to improve the accumulation of a recombinant protein in plant cells require an appreciation of the processes of gene transcription, mRNA stability, processing, and export, and translation initiation and efficiency. Likewise, many post-translational factors must be considered, including protein stability, protein function and activity, and protein targeting. Moreover, we need to understand how the various processes leading from the gene to the functional protein are interdependent and functionally linked. Manipulation of the post-translational processing machinery of plant cells, especially that for N-linked glycosylation and glycan processing, is a challenging and exciting area. The functions of N-glycan heterogeneity and microheterogeneity, especially with respect to protein function, stability, and transport, are poorly understood and this represents an important area of cell biology.

scholarly journals NS1 Recombinant Proteins Are Efficiently Produced in Pichia pastoris and Have Great Potential for Use in Diagnostic Kits for Dengue Virus Infections

Diagnostics ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 379
Author(s):  
Mariana Fonseca Xisto ◽  
John Willians Oliveira Prates ◽  
Ingrid Marques Dias ◽  
Roberto Sousa Dias ◽  
Cynthia Canedo da Silva ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Dengue Virus ◽  
Recombinant Proteins ◽  
Large Scale ◽  
Limiting Factor ◽  
Scale Production ◽  
Global Public Health ◽  
Virus Infections ◽  
Correct Treatment ◽  
Diagnostic Kits

Dengue is one of the major diseases causing global public health concerns. Despite technological advances in vaccine production against all its serotypes, it is estimated that the dengue virus is responsible for approximately 390 million infections per year. Laboratory diagnosis has been the key point for the correct treatment and prevention of this disease. Currently, the limiting factor in the manufacture of dengue diagnostic kits is the large-scale production of the non-structural 1 (NS1) antigen used in the capture of the antibody present in the infected patients’ serum. In this work, we demonstrate the production of the non-structural 1 protein of dengue virus (DENV) serotypes 1–4 (NS1-DENV1, NS1-DENV2, NS1-DENV3, and NS1-DENV4) in the methylotrophic yeast Pichia pastoris KM71H. Secreted recombinant protein was purified by affinity chromatography and characterized by SDS-PAGE and ELISA. The objectives of this study were achieved, and the results showed that P. pastoris is a good heterologous host and worked well in the production of NS1DENV 1–4 recombinant proteins. Easy to grow and quick to obtain, this yeast secreted ready-to-use proteins, with a final yield estimated at 2.8–4.6 milligrams per liter of culture. We reached 85–91% sensitivity and 91–93% specificity using IgM as a target, and for anti-dengue IgG, 83–87% sensitivity and 81–93% specificity were achieved. In this work, we conclude that the NS1 recombinant proteins are efficiently produced in P. pastoris and have great potential for use in diagnostic kits for dengue virus infections. The transformed yeast obtained can be used for production in industrial-scale bioreactors.

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