Transcriptome Analysis Unveils the Effects of Proline on Gene Expression in the Yeast Komagataella phaffii

Komagataella phaffii yeast is one of the most important biocompounds producing microorganisms in modern biotechnology. Optimization of media recipes and cultivation strategies is key to successful synthesis of recombinant proteins. The complex effects of proline on gene expression in the yeast K. phaffii was analyzed on the transcriptome level in this work. Our analysis revealed drastic changes in gene expression when K. phaffii was grown in proline-containing media in comparison to ammonium sulphate-containing media. Around 18.9% of all protein-encoding genes were differentially expressed in the experimental conditions. Proline is catabolized by K. phaffii even in the presence of other nitrogen, carbon and energy sources. This results in the repression of genes involved in the utilization of other element sources, namely methanol. We also found that the repression of AOX1 gene promoter with proline can be partially reversed by the deletion of the KpPUT4.2 gene.

Vaccination against SARS-CoV-2: a human enhancement story

Background Vaccination is an essential strategy for mitigating the COVID-19 pandemic. Besides its significance as a public health measure, vaccination is a sophisticated example of modern biotechnology. Since vaccination gives the human body an ability that it does not naturally possess, the question arises as to its classification as Human Enhancement. Main Body Exemplified on a selection of different definitions, we conclude that vaccinations may indeed be classified and treated as a form of Human Enhancement. This raises some ethical issues that are notorious in the broad field of Human Enhancement. A study with N = 67 participants revealed that vaccinations are perceived neither as a clear nor poor example of Human Enhancement. Conclusion We argue that qualifying vaccination technology as Human Enhancement does not provide convincing arguments to reject vaccination. By examining the Human Enhancement debate and the similarities to the issue of vaccination shown here, policymakers can learn valuable lessons regarding mass vaccination programs’ current and future handling.

KESEDARAN, PENGETAHUAN DAN SIKAP MASYARAKAT AWAM TERHADAP VAKSIN COVID-19 KONVENSIONAL DAN BERASASKAN BIOTEKNOLOGI MODEN DI MALAYSIA (Public Awareness, Knowledge, and Attitude towards Conventional and Modern Biotechnology Based Vaccines in Malaysia)

The COVID-19 pandemic has brought about a paradigm shift in the role and contribution of biotechnology in curbing the spread of SARS-CoV-2 virus worldwide with the production of COVID-19 vaccines either through conventional or modern biotechnology-based methods. Malaysia, which is currently running the National COVID-19 Immunization Program, has approved the conditional registration of several types of vaccines for use during pandemic for its citizens to achieve the targeted level of herd immunity. Thus, this study aims to assess the level of public awareness, self-rated knowledge, and attitude towards three different types of COVID-19 vaccines in Malaysia namely inactivated virus vaccines such as Sinovac, viral vector vaccines such as AstraZeneca and mRNA vaccines such as Pfizer. A survey was conducted using an online survey platform on Malaysians from 19 June to 2 July 2021. Of the 357 respondents, the mean age was 34.75 ± 10.35 with 67.2% of them from STEM-related educational background. Results of the study shows that the Malaysian public were highly aware of the three types of COVID-19 vaccines, but their self-rated knowledge was moderate. Generally, the public were positive towards vaccines. They perceived all the vaccines as having high benefits and highly acceptable from their religious point of view which was translated into high intention to adopt. However, they acknowledged moderate worries on the possible risks of the vaccines. The results of this study play an important role in evaluating the readiness of the Malaysian community to adopt COVID-19 vaccines besides providing an insight into the current attitude of the public towards the vaccination efforts coordinated by the Malaysian government. Keywords: Public awareness; Self-rated knowledge; Behavioural intention; COVID-19 vaccines; Conventional; Modern biotechnology

Von der Wurzel ins Labor: Meerrettichperoxidase produziert in E. coli

The enzyme Horseradish Peroxidase (HRP) is omnipresent in modern biotechnology. Although promising for therapeutic purposes, no suitable production process for this enzyme has been available until now. Medical applications require the enzyme to be highly pure, homogenous and well-defined. We have developed an efficient production process for recombinant HRP from Escherichia coli inclusion bodies. With this strategy we are able to provide active, highly pure and non-glycosylated enzyme at competitive yields.

Modern biotechnology tools and biosecurity in the Middle East

The global health system is under a constant threat from microbial outbreaks. The innovation in genetic engineering has created an existential threat to national, regional and international security. This threat, that can edit microbial or human genomes, requires global attention. In the current review, a comprehensive literature search was conducted using PubMed, SCOPUS and Google Scholar to identify literature discussing modern biotechnology tools as well as relevance to biosafety in the Middle east region. This review was undertaken to provide an overview of biological threats due to advancements in genetic engineering, making it possible to insert or delete specific genes to increase the virulence of particular microbes. These pathogens or other toxic factors can be multiplied by technology, creating new biological weapons. Genome editing technologies including meganucleases (MNs), zinc finger nucleases (ZFNs), transcription activator-like effector (TALE)-nucleases (TALENs) and recently discovered clustered regularly interspaced short palindromic repeats (CRISPR/Cas) induce a double strand break at specific DNA target site. Genome editing technologies lead to an irreversible and permanent alteration of the genetic code and therefore, can inevitably result in security risks. Vulnerabilities in Middle Eastern laboratories raise the prospect of high levels of pathogenic microbes potentially creating a weakness in the diagnosis and monitoring of epidemics. Furthermore, the lack of regional legislation to regulate biosafety and biosecurity may lead to biological threat at the regional level.

Alteration of Flower Color in Viola cornuta cv. “Lutea Splendens” through Metabolic Engineering of Capsanthin/Capsorubin Synthesis

Flower color is an important characteristic that determines the commercial value of ornamental plants. The development of modern biotechnology methods such as genetic engineering enables the creation of new flower colors that cannot be achieved with classical methods of hybridization or mutational breeding. This is the first report on the successful Agrobacterium-mediated genetic transformation of Viola cornuta L. The hypocotyl explants of cv. “Lutea Splendens” variety with yellow flowers were transformed with A. tumefaciens carrying empty pWBVec10a vector (Llccs−) or pWBVec10a/CaMV 35S::Llccs::TNos vector (Llccs+) for capsanthin/capsorubin synthase gene (Llccs) from tiger lily (Lilium lancifolium). A comparative study of shoot multiplication, rooting ability during culture in vitro, as well as phenotypic characteristics of untransformed (control) and transgenic Llccs− and Llccs+ plants during ex vitro growth and flowering is presented. Successful integration of Llccs transgene allows the synthesis of red pigment capsanthin in petal cells that gives flowers different shades of an orange/reddish color. We demonstrate that the ectopic expression of Llccs gene in ornamental plants, such as V. cornuta “Lutea Splendens” could successfully be used to change flower color from yellow to different shades of orange.

BioCAT: PSSM-based algorithm to search biosynthetic gene clusters producing nonribosomal peptides with known structure

Motivation: Nonribosomal peptides (NRPs) are a class of secondary metabolites synthesized by multimodular enzymes named nonribosomal peptide synthetases (NRPSs) and mainly produced by bacteria and fungi. It has been shown that NRPs have a huge structural and functional diversity including antimicrobial activity, therefore, they are of increasing interest for modern biotechnology. Methods such as NMR and LC-MS/MS allow to determine NRP structure precisely, but it is often not a trivial task to find natural producers of them. Today, searches are usually performed manually, mostly with tools such as antiSMASH or Prism. However, there are cases when potential producers should be found among hundreds of strains, for instance, when analyzing metagenomes data. Thus, the development of automated approaches is a high-priority task for further NRP research. Results: We developed BioCAT, a two-side approach to find biosynthesys gene clusters (BGCs) which may produce a given NRP when the structure of interesting NRP has already been found. Formally, the BioCAT unites the antiSMASH software and the rBAN retrosynthesis tool but some improvements were added to both gene cluster and NRP chemical structure analyses. The main feature of the method is PSSM usage to store specificities of NRPS modules, which has increased the alignment quality in comparison with more strict approaches developed earlier. An ensemble model was implemented to calculate the final alignment score. We tested the method on a manually curated NRP producers database and compared it with a competing tool called GARLIC. Finally, we showed the method applicability on a several external examples.

NEXT GENERATION SEQUENCING TECHNOLOGIES TOWARDS EXPLORATION OF MEDICINAL PLANTS

With the ever-increasing population, the plant cover is decreasing at an alarming rate. The medicinal plants are most affected by this because they are present in the last tier of cultivation. Let it be pharmaceutical companies or people using it for herbalism, medicinal plants have been exploited without getting a chance to flourish in their natural environment. Modern biotechnology acts as a bridge between the cultivation and utilization of medicinal plants. Next Generation Sequencing (NGS) technology which is a decade old but emerging field helps to unveil the importance of medicinal plants. Thus, it paves the way for sustenance of medicinal plants by molecular breeding, micropropagation, large-scale tissue culture, and other methods to conserve the plants with great medicinal value. Various NGS technologies can be found in the market like Ilumina, PacBio, Ion Torrent, and others. The present review will summarize the NGS technologies and their potential use to study the genomes, transcriptome, epigenome, and interactome of medicinal plants towards the identification of bioactive compounds.