k lactis protein expression kit  (New England Biolabs)


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    Name:
    K lactisProtein Expression Kit
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    Catalog Number:
    E1000
    Price:
    935
    Category:
    Other Kits
    Applications:
    Proteomics & Glycomics
    Size:
    1 set
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    Structured Review

    New England Biolabs k lactis protein expression kit
    K lactisProtein Expression Kit

    https://www.bioz.com/result/k lactis protein expression kit/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    k lactis protein expression kit - by Bioz Stars, 2021-07
    95/100 stars

    Images

    1) Product Images from "Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin"

    Article Title: Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.03151-13

    Cirsin PSI fused to the α-MF leader sequence is efficiently integrated into the K. lactis genome and secreted into culture medium. (A) Schematic representation of the α-MF_PSI(His) 6 (top) and α-MF_PSI(N86S)(His) 6 (bottom) fusion proteins. Both sequences comprise the α-MF preprodomain sequence and a hexahistidine tag. The two putative N-glycosylation sites in the α-MF leader sequence (NGT and NTT) are indicated. A putative N-glycosylation site (NET) is present in the α-MF_PSI(His) 6 sequence, while in the construct α-MF_PSI(N86S)(His) 6 , this glycosylation site is mutated (N86S). (B) Confirmation by PCR of integration of the expression cassettes into the K. lactis genome. The numbers correspond to the selected clones and “c” to the amplification product using genomic DNA from untransformed K. lactis strain GG799 (negative control). (Top) α-MF_PSI(His) 6 . (Bottom) α-MF_PSI(N86S)(His) 6 . (C) Western blot analysis of recombinant cirsin PSI expression/secretion into culture media with an anti-His antibody. The numbers above the gels correspond to selected clones shown in panel B, and the lane marked with a c corresponds to the analysis of the culture medium of untransformed K. lactis GG799 cells (negative control). (Left) α-MF_PSI(His) 6 transformants. (Right) α-MF_PSI(N86S)(His) 6 transformants.
    Figure Legend Snippet: Cirsin PSI fused to the α-MF leader sequence is efficiently integrated into the K. lactis genome and secreted into culture medium. (A) Schematic representation of the α-MF_PSI(His) 6 (top) and α-MF_PSI(N86S)(His) 6 (bottom) fusion proteins. Both sequences comprise the α-MF preprodomain sequence and a hexahistidine tag. The two putative N-glycosylation sites in the α-MF leader sequence (NGT and NTT) are indicated. A putative N-glycosylation site (NET) is present in the α-MF_PSI(His) 6 sequence, while in the construct α-MF_PSI(N86S)(His) 6 , this glycosylation site is mutated (N86S). (B) Confirmation by PCR of integration of the expression cassettes into the K. lactis genome. The numbers correspond to the selected clones and “c” to the amplification product using genomic DNA from untransformed K. lactis strain GG799 (negative control). (Top) α-MF_PSI(His) 6 . (Bottom) α-MF_PSI(N86S)(His) 6 . (C) Western blot analysis of recombinant cirsin PSI expression/secretion into culture media with an anti-His antibody. The numbers above the gels correspond to selected clones shown in panel B, and the lane marked with a c corresponds to the analysis of the culture medium of untransformed K. lactis GG799 cells (negative control). (Left) α-MF_PSI(His) 6 transformants. (Right) α-MF_PSI(N86S)(His) 6 transformants.

    Techniques Used: Sequencing, Construct, Polymerase Chain Reaction, Expressing, Clone Assay, Amplification, Negative Control, Western Blot, Recombinant

    Recombinant cirsin PSI displays antifungal activity. (A) Confrontation assays between the selected K. lactis transformant secreting wt PSI [SP_PSI(His) 6 ] and three phytopathogenic fungi ( A. alternata , L. infectoria , and D. biseptata ). Untransformed K. lactis (strain GG799) and a K. lactis transformant expressing GFP were used as negative controls. The antifungal activity is given by the percentage of mycelial growth inhibition {MGI = [(free growth of the fungi − growth of the fungi in the presence of yeast)/(free growth of the fungi)] × 100}. All assays were performed in duplicate. (B) Representative plates showing antifungal activity toward A. alternata and L. infectoria . (C and D) Disk diffusion test. Impregnated disks with a purified sample of recombinant nonglycosylated cirsin PSI (0.47 mg/ml) were assayed against the fungus A. alternata by evaluating the appearance of a clear zone of inhibition. The assay was performed in triplicate at two different pH values (pH 5.0 and 6.8) using the antifungal amphotericin B (0.25 mg/ml) as a positive control. (C) Antifungal activity is represented by the percentage of mycelial growth inhibition {MGI = [(growth of the fungi in the presence of sample buffer − growth of the fungi in the presence of PSI)/(growth of fungi in the presence of sample buffer)] × 100}. (D) Representative plates showing antifungal activity of PSI toward A. alternata . The error bars indicate standard deviations.
    Figure Legend Snippet: Recombinant cirsin PSI displays antifungal activity. (A) Confrontation assays between the selected K. lactis transformant secreting wt PSI [SP_PSI(His) 6 ] and three phytopathogenic fungi ( A. alternata , L. infectoria , and D. biseptata ). Untransformed K. lactis (strain GG799) and a K. lactis transformant expressing GFP were used as negative controls. The antifungal activity is given by the percentage of mycelial growth inhibition {MGI = [(free growth of the fungi − growth of the fungi in the presence of yeast)/(free growth of the fungi)] × 100}. All assays were performed in duplicate. (B) Representative plates showing antifungal activity toward A. alternata and L. infectoria . (C and D) Disk diffusion test. Impregnated disks with a purified sample of recombinant nonglycosylated cirsin PSI (0.47 mg/ml) were assayed against the fungus A. alternata by evaluating the appearance of a clear zone of inhibition. The assay was performed in triplicate at two different pH values (pH 5.0 and 6.8) using the antifungal amphotericin B (0.25 mg/ml) as a positive control. (C) Antifungal activity is represented by the percentage of mycelial growth inhibition {MGI = [(growth of the fungi in the presence of sample buffer − growth of the fungi in the presence of PSI)/(growth of fungi in the presence of sample buffer)] × 100}. (D) Representative plates showing antifungal activity of PSI toward A. alternata . The error bars indicate standard deviations.

    Techniques Used: Recombinant, Activity Assay, Expressing, Inhibition, Diffusion-based Assay, Purification, Positive Control

    2) Product Images from "Enhancing the Heterologous Fructosyltransferase Activity of Kluyveromyces lactis: Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing"

    Article Title: Enhancing the Heterologous Fructosyltransferase Activity of Kluyveromyces lactis: Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing

    Journal: Frontiers in Bioengineering and Biotechnology

    doi: 10.3389/fbioe.2020.607507

    Chromatograms showing FOS catalytic products in 100 μL cell-free crude enzyme solution of K. lactis GG799 and GG799Δ Inv with 600 g L −1 sucrose solution for 2 h at 70°C. The peaks are annotated as follows: A = fructose, B = glucose, C = sucrose, D = neokestose, E = 1-kestose, F = 6-kestose, G = nystose, and H = 1 F -fructofranosylnystose.
    Figure Legend Snippet: Chromatograms showing FOS catalytic products in 100 μL cell-free crude enzyme solution of K. lactis GG799 and GG799Δ Inv with 600 g L −1 sucrose solution for 2 h at 70°C. The peaks are annotated as follows: A = fructose, B = glucose, C = sucrose, D = neokestose, E = 1-kestose, F = 6-kestose, G = nystose, and H = 1 F -fructofranosylnystose.

    Techniques Used:

    Time course of the online measured dissolved oxygen (DO) concentration and cell dry weight (CDW) in a fed-batch process using the Labfors3 system. (A) Production strain K. lactis GG799 and (B) mutant strain K. lactis GG799Δ Inv .
    Figure Legend Snippet: Time course of the online measured dissolved oxygen (DO) concentration and cell dry weight (CDW) in a fed-batch process using the Labfors3 system. (A) Production strain K. lactis GG799 and (B) mutant strain K. lactis GG799Δ Inv .

    Techniques Used: Concentration Assay, Mutagenesis

    Related Articles

    Transformation Assay:

    Article Title: Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin
    Article Snippet: After centrifugation, the cells were washed and then resuspended in 1 M sorbitol (cold and sterile) and kept in ice until transformation. .. Before yeast transformation, expression cassettes were obtained by linearizing all pKLAC1-PSI constructs with SacII, according to the manufacturers' instructions ( K. lactis Protein Expression kit; New England BioLabs). .. K. lactis competent cells were then incubated with purified DNA from each expression cassette (2 μg DNA) and transformed using a GenePulser Xcell electroporation system (Bio-Rad) under the following conditions: 1.5 kV, 25 μF, and 200 Ω.

    Expressing:

    Article Title: Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin
    Article Snippet: After centrifugation, the cells were washed and then resuspended in 1 M sorbitol (cold and sterile) and kept in ice until transformation. .. Before yeast transformation, expression cassettes were obtained by linearizing all pKLAC1-PSI constructs with SacII, according to the manufacturers' instructions ( K. lactis Protein Expression kit; New England BioLabs). .. K. lactis competent cells were then incubated with purified DNA from each expression cassette (2 μg DNA) and transformed using a GenePulser Xcell electroporation system (Bio-Rad) under the following conditions: 1.5 kV, 25 μF, and 200 Ω.

    Article Title: Enhancing the Heterologous Fructosyltransferase Activity of Kluyveromyces lactis: Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing
    Article Snippet: We therefore used the CRISPR/Cas9 system to inactivate the native K. lactis invertase gene and tested the biomass yield and transferase activity against the original production strain in batch and fed-batch fermentations. .. K. lactis Production Mutant The K. lactis GG799 production host was generated as previously described (Spohner and Czermak, ) by introducing an FFT gene from A. terreus NIH2624 into the K. lactis GG799 wild-type strain from the K. lactis Protein Expression Kit (New England Biolabs, Frankfurt, Germany). .. CRISPR/Cas9 Plasmid and Invertase Deletion The CRISPR/Cas9 system was used to inactivate the invertase gene (GenBank: AF079370.1 ) in the K. lactis GG799 mutant.

    Article Title: The medicinal plant Andrographis paniculata genome provides insight into biosynthesis of the bioactive diterpenoid neoandrographolide
    Article Snippet: The UGTs were sub-cloned to construct pMAL-UGT expression vectors, which were then transformed into E. coli strain Novablue competent cells. .. To induce protein expression, 0.3 mM of isopropyl β-D-thiogalactoside (IPTG)was added when the OD value of cell culture (grown at 37°C) reached 0.5. .. The MBP-fusion proteins were purified using protocols for the pMAL Protein Fusion and Purification System (New England BioLabs).

    Article Title: Expression of human c-reactive protein in different systems and its purification from Leishmania tarentolae.
    Article Snippet: With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation.

    Article Title: Application of fluorescence correlation spectroscopy to study substrate binding in styrene maleic acid lipid copolymer encapsulated ABCG2
    Article Snippet: The obtained images were manually assessed for relative brightness as an indicator of protein expression level. .. Protein expression was further enhanced by the addition of sodium butyrate (10 mM) to cell cultures 24 h prior to harvesting. .. To facilitate greater cell densities and yields for protein expression, stably transfected His-SNAP-ABCG2-expressing HEK293T cells were adapted to suspension culture by agitation at 180 rpm in flat-bottomed borosilicate vessels with high levels of cell viability ( > 95%) maintained in media (4.5 g.L−1 glucose Dulbecco's modified Eagle medium; DMEM) supplemented with 10% v/v foetal bovine serum (FBS).

    Construct:

    Article Title: Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin
    Article Snippet: After centrifugation, the cells were washed and then resuspended in 1 M sorbitol (cold and sterile) and kept in ice until transformation. .. Before yeast transformation, expression cassettes were obtained by linearizing all pKLAC1-PSI constructs with SacII, according to the manufacturers' instructions ( K. lactis Protein Expression kit; New England BioLabs). .. K. lactis competent cells were then incubated with purified DNA from each expression cassette (2 μg DNA) and transformed using a GenePulser Xcell electroporation system (Bio-Rad) under the following conditions: 1.5 kV, 25 μF, and 200 Ω.

    Mutagenesis:

    Article Title: Enhancing the Heterologous Fructosyltransferase Activity of Kluyveromyces lactis: Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing
    Article Snippet: We therefore used the CRISPR/Cas9 system to inactivate the native K. lactis invertase gene and tested the biomass yield and transferase activity against the original production strain in batch and fed-batch fermentations. .. K. lactis Production Mutant The K. lactis GG799 production host was generated as previously described (Spohner and Czermak, ) by introducing an FFT gene from A. terreus NIH2624 into the K. lactis GG799 wild-type strain from the K. lactis Protein Expression Kit (New England Biolabs, Frankfurt, Germany). .. CRISPR/Cas9 Plasmid and Invertase Deletion The CRISPR/Cas9 system was used to inactivate the invertase gene (GenBank: AF079370.1 ) in the K. lactis GG799 mutant.

    Generated:

    Article Title: Enhancing the Heterologous Fructosyltransferase Activity of Kluyveromyces lactis: Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing
    Article Snippet: We therefore used the CRISPR/Cas9 system to inactivate the native K. lactis invertase gene and tested the biomass yield and transferase activity against the original production strain in batch and fed-batch fermentations. .. K. lactis Production Mutant The K. lactis GG799 production host was generated as previously described (Spohner and Czermak, ) by introducing an FFT gene from A. terreus NIH2624 into the K. lactis GG799 wild-type strain from the K. lactis Protein Expression Kit (New England Biolabs, Frankfurt, Germany). .. CRISPR/Cas9 Plasmid and Invertase Deletion The CRISPR/Cas9 system was used to inactivate the invertase gene (GenBank: AF079370.1 ) in the K. lactis GG799 mutant.

    Cell Culture:

    Article Title: The medicinal plant Andrographis paniculata genome provides insight into biosynthesis of the bioactive diterpenoid neoandrographolide
    Article Snippet: The UGTs were sub-cloned to construct pMAL-UGT expression vectors, which were then transformed into E. coli strain Novablue competent cells. .. To induce protein expression, 0.3 mM of isopropyl β-D-thiogalactoside (IPTG)was added when the OD value of cell culture (grown at 37°C) reached 0.5. .. The MBP-fusion proteins were purified using protocols for the pMAL Protein Fusion and Purification System (New England BioLabs).

    SDS Page:

    Article Title: Expression of human c-reactive protein in different systems and its purification from Leishmania tarentolae.
    Article Snippet: With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation.

    Western Blot:

    Article Title: Expression of human c-reactive protein in different systems and its purification from Leishmania tarentolae.
    Article Snippet: With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation.

    Plasmid Preparation:

    Article Title: Expression of human c-reactive protein in different systems and its purification from Leishmania tarentolae.
    Article Snippet: With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation. .. With its homo-pentameric structure and calcium-dependent specificity for phosphocholine (PCh), human c-reactive protein (CRP) is produced by the liver and secreted in elevated quantities in response to inflammation.

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    New England Biolabs k lactis protein expression kit
    Cirsin <t>PSI</t> fused to the α-MF leader sequence is efficiently integrated into the K. <t>lactis</t> genome and secreted into culture medium. (A) Schematic representation of the α-MF_PSI(His) 6 (top) and α-MF_PSI(N86S)(His) 6 (bottom) fusion proteins. Both sequences comprise the α-MF preprodomain sequence and a hexahistidine tag. The two putative N-glycosylation sites in the α-MF leader sequence (NGT and NTT) are indicated. A putative N-glycosylation site (NET) is present in the α-MF_PSI(His) 6 sequence, while in the construct α-MF_PSI(N86S)(His) 6 , this glycosylation site is mutated (N86S). (B) Confirmation by PCR of integration of the expression cassettes into the K. lactis genome. The numbers correspond to the selected clones and “c” to the amplification product using genomic DNA from untransformed K. lactis strain GG799 (negative control). (Top) α-MF_PSI(His) 6 . (Bottom) α-MF_PSI(N86S)(His) 6 . (C) Western blot analysis of recombinant cirsin PSI expression/secretion into culture media with an anti-His antibody. The numbers above the gels correspond to selected clones shown in panel B, and the lane marked with a c corresponds to the analysis of the culture medium of untransformed K. lactis GG799 cells (negative control). (Left) α-MF_PSI(His) 6 transformants. (Right) α-MF_PSI(N86S)(His) 6 transformants.
    K Lactis Protein Expression Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/k lactis protein expression kit/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    k lactis protein expression kit - by Bioz Stars, 2021-07
    95/100 stars
      Buy from Supplier

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    Cirsin PSI fused to the α-MF leader sequence is efficiently integrated into the K. lactis genome and secreted into culture medium. (A) Schematic representation of the α-MF_PSI(His) 6 (top) and α-MF_PSI(N86S)(His) 6 (bottom) fusion proteins. Both sequences comprise the α-MF preprodomain sequence and a hexahistidine tag. The two putative N-glycosylation sites in the α-MF leader sequence (NGT and NTT) are indicated. A putative N-glycosylation site (NET) is present in the α-MF_PSI(His) 6 sequence, while in the construct α-MF_PSI(N86S)(His) 6 , this glycosylation site is mutated (N86S). (B) Confirmation by PCR of integration of the expression cassettes into the K. lactis genome. The numbers correspond to the selected clones and “c” to the amplification product using genomic DNA from untransformed K. lactis strain GG799 (negative control). (Top) α-MF_PSI(His) 6 . (Bottom) α-MF_PSI(N86S)(His) 6 . (C) Western blot analysis of recombinant cirsin PSI expression/secretion into culture media with an anti-His antibody. The numbers above the gels correspond to selected clones shown in panel B, and the lane marked with a c corresponds to the analysis of the culture medium of untransformed K. lactis GG799 cells (negative control). (Left) α-MF_PSI(His) 6 transformants. (Right) α-MF_PSI(N86S)(His) 6 transformants.

    Journal: Applied and Environmental Microbiology

    Article Title: Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin

    doi: 10.1128/AEM.03151-13

    Figure Lengend Snippet: Cirsin PSI fused to the α-MF leader sequence is efficiently integrated into the K. lactis genome and secreted into culture medium. (A) Schematic representation of the α-MF_PSI(His) 6 (top) and α-MF_PSI(N86S)(His) 6 (bottom) fusion proteins. Both sequences comprise the α-MF preprodomain sequence and a hexahistidine tag. The two putative N-glycosylation sites in the α-MF leader sequence (NGT and NTT) are indicated. A putative N-glycosylation site (NET) is present in the α-MF_PSI(His) 6 sequence, while in the construct α-MF_PSI(N86S)(His) 6 , this glycosylation site is mutated (N86S). (B) Confirmation by PCR of integration of the expression cassettes into the K. lactis genome. The numbers correspond to the selected clones and “c” to the amplification product using genomic DNA from untransformed K. lactis strain GG799 (negative control). (Top) α-MF_PSI(His) 6 . (Bottom) α-MF_PSI(N86S)(His) 6 . (C) Western blot analysis of recombinant cirsin PSI expression/secretion into culture media with an anti-His antibody. The numbers above the gels correspond to selected clones shown in panel B, and the lane marked with a c corresponds to the analysis of the culture medium of untransformed K. lactis GG799 cells (negative control). (Left) α-MF_PSI(His) 6 transformants. (Right) α-MF_PSI(N86S)(His) 6 transformants.

    Article Snippet: Before yeast transformation, expression cassettes were obtained by linearizing all pKLAC1-PSI constructs with SacII, according to the manufacturers' instructions ( K. lactis Protein Expression kit; New England BioLabs).

    Techniques: Sequencing, Construct, Polymerase Chain Reaction, Expressing, Clone Assay, Amplification, Negative Control, Western Blot, Recombinant

    Recombinant cirsin PSI displays antifungal activity. (A) Confrontation assays between the selected K. lactis transformant secreting wt PSI [SP_PSI(His) 6 ] and three phytopathogenic fungi ( A. alternata , L. infectoria , and D. biseptata ). Untransformed K. lactis (strain GG799) and a K. lactis transformant expressing GFP were used as negative controls. The antifungal activity is given by the percentage of mycelial growth inhibition {MGI = [(free growth of the fungi − growth of the fungi in the presence of yeast)/(free growth of the fungi)] × 100}. All assays were performed in duplicate. (B) Representative plates showing antifungal activity toward A. alternata and L. infectoria . (C and D) Disk diffusion test. Impregnated disks with a purified sample of recombinant nonglycosylated cirsin PSI (0.47 mg/ml) were assayed against the fungus A. alternata by evaluating the appearance of a clear zone of inhibition. The assay was performed in triplicate at two different pH values (pH 5.0 and 6.8) using the antifungal amphotericin B (0.25 mg/ml) as a positive control. (C) Antifungal activity is represented by the percentage of mycelial growth inhibition {MGI = [(growth of the fungi in the presence of sample buffer − growth of the fungi in the presence of PSI)/(growth of fungi in the presence of sample buffer)] × 100}. (D) Representative plates showing antifungal activity of PSI toward A. alternata . The error bars indicate standard deviations.

    Journal: Applied and Environmental Microbiology

    Article Title: Establishing the Yeast Kluyveromyces lactis as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin

    doi: 10.1128/AEM.03151-13

    Figure Lengend Snippet: Recombinant cirsin PSI displays antifungal activity. (A) Confrontation assays between the selected K. lactis transformant secreting wt PSI [SP_PSI(His) 6 ] and three phytopathogenic fungi ( A. alternata , L. infectoria , and D. biseptata ). Untransformed K. lactis (strain GG799) and a K. lactis transformant expressing GFP were used as negative controls. The antifungal activity is given by the percentage of mycelial growth inhibition {MGI = [(free growth of the fungi − growth of the fungi in the presence of yeast)/(free growth of the fungi)] × 100}. All assays were performed in duplicate. (B) Representative plates showing antifungal activity toward A. alternata and L. infectoria . (C and D) Disk diffusion test. Impregnated disks with a purified sample of recombinant nonglycosylated cirsin PSI (0.47 mg/ml) were assayed against the fungus A. alternata by evaluating the appearance of a clear zone of inhibition. The assay was performed in triplicate at two different pH values (pH 5.0 and 6.8) using the antifungal amphotericin B (0.25 mg/ml) as a positive control. (C) Antifungal activity is represented by the percentage of mycelial growth inhibition {MGI = [(growth of the fungi in the presence of sample buffer − growth of the fungi in the presence of PSI)/(growth of fungi in the presence of sample buffer)] × 100}. (D) Representative plates showing antifungal activity of PSI toward A. alternata . The error bars indicate standard deviations.

    Article Snippet: Before yeast transformation, expression cassettes were obtained by linearizing all pKLAC1-PSI constructs with SacII, according to the manufacturers' instructions ( K. lactis Protein Expression kit; New England BioLabs).

    Techniques: Recombinant, Activity Assay, Expressing, Inhibition, Diffusion-based Assay, Purification, Positive Control

    Chromatograms showing FOS catalytic products in 100 μL cell-free crude enzyme solution of K. lactis GG799 and GG799Δ Inv with 600 g L −1 sucrose solution for 2 h at 70°C. The peaks are annotated as follows: A = fructose, B = glucose, C = sucrose, D = neokestose, E = 1-kestose, F = 6-kestose, G = nystose, and H = 1 F -fructofranosylnystose.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Enhancing the Heterologous Fructosyltransferase Activity of Kluyveromyces lactis: Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing

    doi: 10.3389/fbioe.2020.607507

    Figure Lengend Snippet: Chromatograms showing FOS catalytic products in 100 μL cell-free crude enzyme solution of K. lactis GG799 and GG799Δ Inv with 600 g L −1 sucrose solution for 2 h at 70°C. The peaks are annotated as follows: A = fructose, B = glucose, C = sucrose, D = neokestose, E = 1-kestose, F = 6-kestose, G = nystose, and H = 1 F -fructofranosylnystose.

    Article Snippet: K. lactis Production Mutant The K. lactis GG799 production host was generated as previously described (Spohner and Czermak, ) by introducing an FFT gene from A. terreus NIH2624 into the K. lactis GG799 wild-type strain from the K. lactis Protein Expression Kit (New England Biolabs, Frankfurt, Germany).

    Techniques:

    Time course of the online measured dissolved oxygen (DO) concentration and cell dry weight (CDW) in a fed-batch process using the Labfors3 system. (A) Production strain K. lactis GG799 and (B) mutant strain K. lactis GG799Δ Inv .

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Enhancing the Heterologous Fructosyltransferase Activity of Kluyveromyces lactis: Developing a Scaled-Up Process and Abolishing Invertase by CRISPR/Cas9 Genome Editing

    doi: 10.3389/fbioe.2020.607507

    Figure Lengend Snippet: Time course of the online measured dissolved oxygen (DO) concentration and cell dry weight (CDW) in a fed-batch process using the Labfors3 system. (A) Production strain K. lactis GG799 and (B) mutant strain K. lactis GG799Δ Inv .

    Article Snippet: K. lactis Production Mutant The K. lactis GG799 production host was generated as previously described (Spohner and Czermak, ) by introducing an FFT gene from A. terreus NIH2624 into the K. lactis GG799 wild-type strain from the K. lactis Protein Expression Kit (New England Biolabs, Frankfurt, Germany).

    Techniques: Concentration Assay, Mutagenesis