scholarly journals Genome Editing to Generate Sake Yeast Strains with Eight Mutations That Confer Excellent Brewing Characteristics

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1299
Author(s):  
Tomoya Chadani ◽  
Shinsuke Ohnuki ◽  
Atsuko Isogai ◽  
Tetsuya Goshima ◽  
Mao Kashima ◽  
...  
Keyword(s):  
Genome Editing ◽  
Small Scale ◽  
Evidence Based ◽  
Considerable Effort ◽  
Recessive Mutations ◽  
Yeast Strains ◽  
Unpleasant Odor ◽  
Genetic Perturbations ◽  
Foam Forming

Sake yeast is mostly diploid, so the introduction of recessive mutations to improve brewing characteristics requires considerable effort. To construct sake yeast with multiple excellent brewing characteristics, we used an evidence-based approach that exploits genome editing technology. Our breeding targeted the AWA1, CAR1, MDE1, and FAS2 genes. We introduced eight mutations into standard sake yeast to construct a non-foam-forming strain that makes sake without producing carcinogens or an unpleasant odor, while producing a sweet ginjo aroma. Small-scale fermentation tests showed that the desired sake could be brewed with our genome-edited strains. The existence of a few unexpected genetic perturbations introduced during breeding proved that genome editing technology is extremely effective for the serial breeding of sake yeast.

scholarly journals Bioreactor Co-Cultivation of High Lipid and Carotenoid Producing Yeast Rhodotorula kratochvilovae and Several Microalgae under Stress

2021 ◽  
Vol 9 (6) ◽  
pp. 1160
Author(s):  
Martin Szotkowski ◽  
Jiří Holub ◽  
Samuel Šimanský ◽  
Klára Hubačová ◽  
Pavlína Sikorová ◽  
...  
Keyword(s):  
Positive Impact ◽  
Small Scale ◽  
Biomass Growth ◽  
Media Composition ◽  
Yeast Strains ◽  
High Lipid ◽  
Algae Growth ◽  
Chlorophyll Production ◽  
Lipid Metabolites ◽  
Present Pilot Study

The co-cultivation of red yeasts and microalgae works with the idea of the natural transport of gases. The microalgae produce oxygen, which stimulates yeast growth, while CO2 produced by yeast is beneficial for algae growth. Both microorganisms can then produce lipids. The present pilot study aimed to evaluate the ability of selected microalgae and carotenogenic yeast strains to grow and metabolize in co-culture. The effect of media composition on growth and metabolic activity of red yeast strains was assessed simultaneously with microalgae mixotrophy. Cultivation was transferred from small-scale co-cultivation in Erlenmeyer flasks to aerated bottles with different inoculation ratios and, finally, to a 3L bioreactor. Among red yeasts, the strain R. kratochvilovae CCY 20-2-26 was selected because of the highest biomass production on BBM medium. Glycerol is a more suitable carbon source in the BBM medium and urea was proposed as a compromise. From the tested microalgae, Desmodesmus sp. were found as the most suitable for co-cultivations with R. kratochvilovae. In all co-cultures, linear biomass growth was found (144 h), and the yield was in the range of 8.78–11.12 g/L of dry biomass. Lipids increased to a final value of 29.62–31.61%. The FA profile was quite stable with the UFA portion at about 80%. Around 1.98–2.49 mg/g CDW of carotenoids with torularhodine as the major pigment were produced, ubiquinone production reached 5.41–6.09 mg/g, and ergosterol yield was 6.69 mg/g. Chlorophyll production was very low at 2.11 mg/g. Pilot experiments have confirmed that carotenogenic yeasts and microalgae are capable of symbiotic co-existence with a positive impact om biomass growth and lipid metabolites yields.

What does it Take to Scale up and Sustain Evidence-Based Practices?

2013 ◽  
Vol 79 (2) ◽  
pp. 195-211 ◽  
Author(s):  
Janette K. Klingner ◽  
Alison G. Boardman ◽  
Kristen L. Mcmaster
Keyword(s):  
Research Policy ◽  
Scale Up ◽  
Scaling Up ◽  
District Leadership ◽  
Small Scale ◽  
Evidence Based ◽  
Policy And Practice ◽  
Evidence Based Practices ◽  
Strategic Reading ◽  
Collaborative Strategic Reading

This article discusses the strategic scaling up of evidence-based practices. The authors draw from the scholarly work of fellow special education researchers and from the field of learning sciences. The article defines scaling up as the process by which researchers or educators initially implement interventions on a small scale, validate them, and then implement them more widely in real-world conditions. Examples of scale-up research are included. The authors discuss challenges to scaling up and sustaining evidence-based practices, followed by factors that can potentially support scaling up, including professional development and district leadership. A case example describes how these issues can play out by highlighting experiences with a Collaborative Strategic Reading (CSR) scale-up research project in a large urban school district. The article concludes by offering recommendations for research, policy, and practice.

scholarly journals Influence of media composition and temperature on volatile aroma production by various wine yeast strains

2008 ◽  
Vol 26 (No. 5) ◽  
pp. 376-382 ◽  
Author(s):  
V. Petravić Tominac ◽  
K. Kovačević Ganić ◽  
D. Komes ◽  
L. Gracin ◽  
M. Banović ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Synthetic Medium ◽  
Aroma Compounds ◽  
Small Scale ◽  
Wine Yeasts ◽  
Wine Production ◽  
Yeast Strains ◽  
Aroma Production ◽  
Volatile Aroma ◽  
Volatile Aroma Compounds

Volatile aroma compounds production by two autochthonous <I>Saccharomyces cerevisiae</I> strains, isolated from Istria region, and three other yeast strains (<I>Saccharomyces bayanus</I> and two commercial <I>Saccharomyces cerevisiae</I> wine yeasts) was investigated on a small scale using synthetic VP4 medium and Graševina must at 12 and 20°C. The results obtained by gas chromatography analyses were compared with the aroma production properties of the native microflora, remaining after Graševina must sulphiting. In both media and at both temperatures, the wine yeasts investigated showed different metabolic profiles regarding the tested volatile aroma compounds, which should be taken in consideration for autochthonous wine production. Although the synthetic medium proved to be appropriate for the investigation of the fermentative properties, the determination of secondary aroma production by wine yeasts has to be conducted by must fermentation or possibly by fermentation of another synthetic medium whose composition would be more similar to must.

scholarly journals Isolation of Auxotrophic Mutants of Diploid Industrial Yeast Strains after UV Mutagenesis

2005 ◽  
Vol 71 (1) ◽  
pp. 312-319 ◽  
Author(s):  
Shinji Hashimoto ◽  
Mayumi Ogura ◽  
Kazuo Aritomi ◽  
Hisashi Hoshida ◽  
Yoshinori Nishizawa ◽  
...  
Keyword(s):  
Selection Procedure ◽  
Isoamyl Acetate ◽  
Gene Replacement ◽  
Industrial Yeast ◽  
Uv Mutagenesis ◽  
Mutant Selection ◽  
Auxotrophic Mutants ◽  
Recessive Mutations ◽  
Alcohol Acetyltransferase ◽  
Yeast Strains

ABSTRACT Auxotrophic mutants of the yeast Saccharomyces cerevisiae are usually isolated in haploid strains because the isolation of recessive mutations in diploids is thought to be difficult due to the presence of two sets of genes. We show here that auxotrophic mutants of diploid industrial sake yeast strains were routinely obtained by a standard mutant selection procedure following UV mutagenesis. We isolated His−, Met−, Lys−, Trp−, Leu−, Arg−, and Ura− auxotrophic mutants of five sake strains, Kyokai no. 7, no. 9, no. 10, no. 701, and no. 901, by screening only 1,700 to 3,400 colonies from each treated strain. Wild-type alleles were cloned and used as markers for transformation. With HIS3 as a selectable marker, the yeast TDH3 overexpression promoter was inserted upstream of ATF1, encoding alcohol acetyltransferase, by one-step gene replacement in a his3 mutant of Kyokai no. 7. The resulting strain contained exclusively yeast DNA, making it acceptable for commercial use, and produced a larger amount of isoamyl acetate, a banana-like flavor. We argue that the generally recognized difficulty of isolating auxotrophic mutants of diploid industrial yeast strains is misleading and that genetic techniques used for haploid laboratory strains are applicable for this purpose.

scholarly journals Postembryonic screen for mutations affecting spine development in zebrafish

2020 ◽  
Author(s):  
Ryan S. Gray ◽  
Roberto Gonzalez ◽  
Sarah D. Ackerman ◽  
Ryoko Minowa ◽  
Johanna F. Griest ◽  
...  
Keyword(s):  
Whole Body ◽  
Small Scale ◽  
Healthy Children ◽  
Synonymous Mutations ◽  
Structural Support ◽  
Recessive Mutations ◽  
Phenotypic Group ◽  
Spine Development ◽  
Lateral Curvature ◽  
Adult Spine

AbstractThe spinal vertebral column gives structural support for the adult body plan, protects the spinal cord, and provides muscle attachment and stability, which allows the animal to move within its environment. The development and maturation of the spine and its physiology involve the integration of multiple musculoskeletal tissues including bone, cartilage, and fibrocartilaginous joints, as well as innervation and control by the nervous system. One of the most common disorders of the spine in human is adolescent idiopathic scoliosis (AIS), which is characterized by the onset of an abnormal lateral curvature of the spine of <10° around adolescence, in otherwise healthy children. The genetic basis of AIS is largely unknown. Systematic genome-wide mutagenesis screens for embryonic phenotypes in zebrafish have been instrumental in the understanding of early patterning of embryonic tissues necessary to build and pattern the embryonic spine. However, the mechanisms required for postembryonic maturation and homeostasis of the spine remain poorly understood. Here we report the results from a small-scale forward genetic screen for adult-viable recessive and dominant mutant zebrafish, displaying overt morphological abnormalities of the adult spine. Germline mutations induced with N-ethyl N-nitrosourea (ENU) were transmitted and screened for dominant phenotypes in 1,229 F1 animals, and subsequently bred to homozygosity in F3 families, from these, 314 haploid genomes were screened for recessive phenotypes. We cumulatively found 39 adult-viable (3 dominant and 36 recessive) mutations each leading to a defect in the morphogenesis of the spine. The largest phenotypic group displayed larval onset axial curvatures, leading to whole-body scoliosis without vertebral dysplasia in adult fish. Pairwise complementation testing within this phenotypic group revealed at least 16 independent mutant loci. Using massively-parallel whole genome or whole exome sequencing and meiotic mapping we defined the molecular identity of several loci for larval onset whole-body scoliosis in zebrafish. We identified a new mutation in the skolios/kinesin family member 6 (kif6) gene, causing neurodevelopmental and ependymal cilia defects in mouse and zebrafish. We also report several recessive alleles of the scospondin and a disintegrin and metalloproteinase with thrombospondin motifs 9 (adamts9) genes, which all display defects in spine morphogenesis. Many of the alleles characterized thus far are non-synonymous mutations in known essential scospondin and adamts9 genes. Our results provide evidence of monogenic traits that are critical for normal spine development in zebrafish, that may help to establish new candidate risk loci for spine disorders in humans.

scholarly journals Evidence-Based Practice (EBP) in Rehabilitative Physiotherapy

2012 ◽  
Author(s):  
Chris Cimdi
Keyword(s):  
Professional Development ◽  
Survey Methodology ◽  
Clinical Decision Making ◽  
Evidence Based Practice ◽  
Research Literature ◽  
Small Scale ◽  
Successful Implementation ◽  
Evidence Based ◽  
Positive Effects ◽  
Wide Range

Evidence-Based Practice (EBP) is considered to be an essential component of clinical service delivery in healthcare. Australian physiotherapists have access to a wide range of resources to support the implementation of EBP. However, research indicates variability in how physiotherapists apply EBP with a range of factors affecting the successful implementation of EBP into everyday clinical practice. Using survey methodology, this study examined whether participation in an EBP professional development workshop is an effective strategy to enhance knowledge, attitudes and behaviours associated with EBP of physiotherapists working in a rehabilitation setting. Results revealed that nearly one third of participants rated their EBP knowledge and skills at a higher level following attendance at the workshop. An increase in the frequency in EBP related activities by physiotherapists including conducting searches and reading or reviewing research literature to support clinical decision-making was observed following attendance at the EBP workshop. Participants’ opinions of the value of EBP in improving the quality of patient care were noted to be more positive following the workshop. This small-scale study provides some promising results regarding the positive effects that designated EBP professional development can have on knowledge, attitudes, and behaviours of physiotherapists in their application of EBP. The elements that are required to further support and sustain a strong EBP culture amongst physiotherapists working in rehabilitation are also discussed.

Middle leading small-scale school projects

2019 ◽  
Vol 33 (7) ◽  
pp. 1733-1745
Author(s):  
Peter Grootenboer ◽  
Kevin Larkin
Keyword(s):  
School Leadership ◽  
Teaching And Learning ◽  
Educational Development ◽  
Single Case ◽  
Small Scale ◽  
Evidence Based ◽  
Case Study Methodology ◽  
Content Type ◽  
School Based ◽  
Middle Leaders

Purpose The authors argue that middle leaders are the key educators in school-based educational development. Schools often secure small-scale funding to engage in government or systemic initiatives, and these projects require a leadership “close to the classroom” if they are to realise sustainable educational gains. This leadership often comes from the middle leaders – those who practice their leading in and around classrooms. The paper aims to discuss this issue. Design/methodology/approach A single case study methodology is used to investigate two middle leaders, leading a small-scale project. Their leading practices are examined using the “theory of practice architectures”, to identify how these practices were enacted within their educational context. Findings While principals play a crucial role in enacting change, it is the middle leaders who are closer to the classroom than most principals, and whose practices more directly impact teaching and learning as they are best placed to ensure that meagre resources are well used to improve student learning. They do this by ensuring that development is collegial and a response to evidence-based needs. Practical implications First, middle leaders need support in facilitating educational development. Second, their leading practice is crucial for sustainable school-based development. Third, site-based educational development occurs most effectively when it is evidence-based. Finally, this form of educational development requires high-level collegiality. Originality/value This paper is original in two key ways: first, it addresses the under-researched practices of middle leaders; and, second it employs the practice theory to understand school leadership and development.

scholarly journals Lager-brewing yeasts in the era of modern genetics

2019 ◽  
Vol 19 (7) ◽  
Author(s):  
Arthur R Gorter de Vries ◽  
Jack T Pronk ◽  
Jean-Marc G Daran
Keyword(s):  
Genome Editing ◽  
Industrial Application ◽  
Strain Improvement ◽  
Brewing Yeast ◽  
Yeast Strains ◽  
Worldwide Production ◽  
Saccharomyces Pastorianus ◽  
Saccharomyces Eubayanus ◽  
Brewing Yeasts

ABSTRACT The yeast Saccharomyces pastorianus is responsible for the annual worldwide production of almost 200 billion liters of lager-type beer. S. pastorianus is a hybrid of Saccharomyces cerevisiae and Saccharomyces eubayanus that has been studied for well over a century. Scientific interest in S. pastorianus intensified upon the discovery, in 2011, of its S. eubayanus ancestor. Moreover, advances in whole-genome sequencing and genome editing now enable deeper exploration of the complex hybrid and aneuploid genome architectures of S. pastorianus strains. These developments not only provide novel insights into the emergence and domestication of S. pastorianus but also generate new opportunities for its industrial application. This review paper combines historical, technical and socioeconomic perspectives to analyze the evolutionary origin and genetics of S. pastorianus. In addition, it provides an overview of available methods for industrial strain improvement and an outlook on future industrial application of lager-brewing yeasts. Particular attention is given to the ongoing debate on whether current S. pastorianus originates from a single or multiple hybridization events and to the potential role of genome editing in developing industrial brewing yeast strains.

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