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Article ; Online: Unnatural amino acids: production and biotechnological potential.

Narancic, Tanja / Almahboub, Sarah A / O'Connor, Kevin E

World journal of microbiology & biotechnology

2019 Volume 35, Issue 4, Page(s) 67

Abstract: Unnatural amino acids (UAAs) are valuable building blocks in the manufacture of a wide range of pharmaceuticals. UAAs exhibit biological activity as free acids and they can be incorporated into linear or cyclic peptides with biological activity. However, ...

Abstract	Unnatural amino acids (UAAs) are valuable building blocks in the manufacture of a wide range of pharmaceuticals. UAAs exhibit biological activity as free acids and they can be incorporated into linear or cyclic peptides with biological activity. However, the scope of biotechnological application of UAAs goes beyond this, as they can be used to investigate the structure and dynamics of proteins, to study protein interactions, or to modulate the activity of proteins in living cells. The means to expand nature's repertoire of amino acids include chemical and biological routes. An UAA can be made through chemical modifications of natural amino acids, or related compounds. These modifications typically rely on utilisation of ligands and palladium catalysts. Employing biocatalysts in the synthesis of UAAs can also afford novel molecules with different physical and chemical properties. A number of transaminases for example have been identified and employed in the production of UAAs. This review will compare the chemical and biological routes for the synthesis of UAAs and provide an overview of their applications.
MeSH term(s)	Amino Acids/biosynthesis ; Amino Acids/chemical synthesis ; Amino Acids/chemistry ; Biocatalysis ; Biotechnology/methods ; Enzymes/metabolism ; Metabolic Engineering ; Protein Engineering/methods
Chemical Substances	Amino Acids ; Enzymes
Language	English
Publishing date	2019-04-08
Publishing country	Germany
Document type	Journal Article ; Review
ZDB-ID	1499109-3
ISSN	1573-0972 ; 0959-3993
ISSN (online)	1573-0972
ISSN	0959-3993
DOI	10.1007/s11274-019-2642-9
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Article: Evaluation of Neutralizing Antibodies Against Highly Pathogenic Coronaviruses: A Detailed Protocol for a Rapid Evaluation of Neutralizing Antibodies Using Vesicular Stomatitis Virus Pseudovirus-Based Assay.

Almahboub, Sarah A / Algaissi, Abdullah / Alfaleh, Mohamed A / ElAssouli, M-Zaki / Hashem, Anwar M

Frontiers in microbiology

2020 Volume 11, Page(s) 2020

Abstract: Emerging highly pathogenic human coronaviruses (CoVs) represent a serious ongoing threat to the public health worldwide. The spike (S) proteins of CoVs are surface glycoproteins that facilitate viral entry into host cells via attachment to their ... ...

Abstract	Emerging highly pathogenic human coronaviruses (CoVs) represent a serious ongoing threat to the public health worldwide. The spike (S) proteins of CoVs are surface glycoproteins that facilitate viral entry into host cells via attachment to their respective cellular receptors. The S protein is believed to be a major immunogenic component of CoVs and a target for neutralizing antibodies (nAbs) and most candidate vaccines. Development of a safe and convenient assay is thus urgently needed to determine the prevalence of CoVs nAbs in the population, to study immune response in infected individuals, and to aid in vaccines and viral entry inhibitor evaluation. While live virus-based neutralization assays are used as gold standard serological methods to detect and measure nAbs, handling of highly pathogenic live CoVs requires strict bio-containment conditions in biosafety level-3 (BSL-3) laboratories. On the other hand, use of replication-incompetent pseudoviruses bearing CoVs S proteins could represent a safe and useful method to detect nAbs in serum samples under biosafety level-2 (BSL-2) conditions. Here, we describe a detailed protocol of a safe and convenient assay to generate vesicular stomatitis virus (VSV)-based pseudoviruses to evaluate and measure nAbs against highly pathogenic CoVs. The protocol covers methods to produce VSV pseudovirus bearing the S protein of the Middle East respiratory syndrome-CoV (MERS-CoV) and the severe acute respiratory syndrome-CoV-2 (SARS-CoV-2), pseudovirus titration, and pseudovirus neutralization assay. Such assay could be adapted by different laboratories and researchers working on highly pathogenic CoVs without the need to handle live viruses in the BSL-3 environment.
Keywords	covid19
Language	English
Publishing date	2020-09-04
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2020.02020
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Construction of VSVΔ51M oncolytic virus expressing human interleukin-12.

Abdulal, Rwaa H / Malki, Jana S / Ghazal, Ezdehar / Alsaieedi, Ahdab A / Almahboub, Sarah A / Khan, Muhammad Yasir / Alsulaiman, Reem M / Ghaith, Mazen M / Abujamel, Turki S / Ganash, Magdah / Mahmoud, Ahmad Bakur / Alkayyal, Almohanad A / Hashem, Anwar M

Frontiers in molecular biosciences

2023 Volume 10, Page(s) 1190669

Abstract: The use of oncolytic viruses (OVs) in combination with cytokines, such as IL-12, is a promising approach for cancer treatment that addresses the limitations of current standard treatments and traditional cancer immunotherapies. IL-12, a proinflammatory ... ...

Abstract	The use of oncolytic viruses (OVs) in combination with cytokines, such as IL-12, is a promising approach for cancer treatment that addresses the limitations of current standard treatments and traditional cancer immunotherapies. IL-12, a proinflammatory cytokine, triggers intracellular signaling pathways that lead to increased apoptosis of tumor cells and enhanced antitumor activity of immune cells via IFN-γ induction, making this cytokine a promising candidate for cancer therapy. Targeted expression of IL-12 within tumors has been shown to play a crucial role in tumor eradication. The recent development of oncolytic viruses enables targeted delivery and expression of IL-12 at the tumor site, thereby addressing the systemic toxicities associated with traditional cancer therapy. In this study, we constructed an oncolytic virus, VSVΔ51M, based on the commercially available VSV wild-type backbone and further modified it to express human IL-12. Our preclinical data confirmed the safety and limited toxicity of the modified virus, VSV-Δ51M-hIL-12, supporting its potential use for clinical development.
Language	English
Publishing date	2023-05-15
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2814330-9
ISSN	2296-889X
ISSN	2296-889X
DOI	10.3389/fmolb.2023.1190669
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Single point mutations reveal amino acid residues important for Chromobacterium violaceum transaminase activity in the production of unnatural amino acids.

Almahboub, Sarah A / Narancic, Tanja / Fayne, Darren / O'Connor, Kevin E

Scientific reports

2018 Volume 8, Issue 1, Page(s) 17397

Abstract: Unnatural amino acids (UAAs) are chiral amines with high application potential in drug discovery and synthesis of other valuable chemicals. Biocatalysis offers the possibility to synthesise novel optically pure UAAs with different physical and chemical ... ...

Abstract	Unnatural amino acids (UAAs) are chiral amines with high application potential in drug discovery and synthesis of other valuable chemicals. Biocatalysis offers the possibility to synthesise novel optically pure UAAs with different physical and chemical properties. While the biocatalytic potential of transaminases in the synthesis of UAAs has been demonstrated, there is still a need to improve the activity with non-native substrates and to understand which amino acids residues are important for activity with these UAAs. Using a rational design approach, six variants of Chromobacterium violaceum DSM30191 transaminase (CV_TA) carrying a single and one variant carrying two substitutions were generated. Among the variants with a single substitution, CV_Y168F showed a 2 to 2.6-fold increased affinity for 2-oxooctanoic acid (2-OOA) and 3-oxobutyric acid (3-OBA) methyl ester used to synthesise an α- and β-UAA. Analysis of the first half of the transaminase reaction showed no change in the activity with the donor (S)-1-phenylethylamine. The combination of W60C and Y168F substitutions improved the CV_TA affinity for 2-OOA 10-fold compared to the wild type. Other substitutions showed no change, or reduced activity with the tested substrates. Our findings provide structural information on CV_TA and demonstrate the potential of rational design for biosynthesis of UAAs.
MeSH term(s)	Amino Acids/genetics ; Biocatalysis ; Chromobacterium/genetics ; Phenethylamines/metabolism ; Point Mutation/genetics ; Transaminases/genetics
Chemical Substances	Amino Acids ; Phenethylamines ; Transaminases (EC 2.6.1.-)
Language	English
Publishing date	2018-11-26
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-018-35688-7
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Article: Evaluation of Neutralizing Antibodies Against Highly Pathogenic Coronaviruses: A Detailed Protocol for a Rapid Evaluation of Neutralizing Antibodies Using Vesicular Stomatitis Virus Pseudovirus-Based Assay

Almahboub, Sarah A. / Algaissi, Abdullah / Alfaleh, Mohamed A. / ElAssouli, M-Zaki / Hashem, Anwar M.

Front. Microbiol.

Abstract	Emerging highly pathogenic human coronaviruses (CoVs) represent a serious ongoing threat to the public health worldwide. The spike (S) proteins of CoVs are surface glycoproteins that facilitate viral entry into host cells via attachment to their respective cellular receptors. The S protein is believed to be a major immunogenic component of CoVs and a target for neutralizing antibodies (nAbs) and most candidate vaccines. Development of a safe and convenient assay is thus urgently needed to determine the prevalence of CoVs nAbs in the population, to study immune response in infected individuals, and to aid in vaccines and viral entry inhibitor evaluation. While live virus-based neutralization assays are used as gold standard serological methods to detect and measure nAbs, handling of highly pathogenic live CoVs requires strict bio-containment conditions in biosafety level-3 (BSL-3) laboratories. On the other hand, use of replication-incompetent pseudoviruses bearing CoVs S proteins could represent a safe and useful method to detect nAbs in serum samples under biosafety level-2 (BSL-2) conditions. Here, we describe a detailed protocol of a safe and convenient assay to generate vesicular stomatitis virus (VSV)-based pseudoviruses to evaluate and measure nAbs against highly pathogenic CoVs. The protocol covers methods to produce VSV pseudovirus bearing the S protein of the Middle East respiratory syndrome-CoV (MERS-CoV) and the severe acute respiratory syndrome-CoV-2 (SARS-CoV-2), pseudovirus titration, and pseudovirus neutralization assay. Such assay could be adapted by different laboratories and researchers working on highly pathogenic CoVs without the need to handle live viruses in the BSL-3 environment.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #798886
Database	COVID19

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Article ; Online: ACE2-Fc and DPP4-Fc decoy receptors against SARS-CoV-2 and MERS-CoV variants: a quick therapeutic option for current and future coronaviruses outbreaks.

Alfaleh, Mohamed A / Alsulaiman, Reem M / Almahboub, Sarah A / Nezamuldeen, Leena / Zawawi, Ayat / Aljehani, Najwa D / Yasir, Muhammad / Abdulal, Rwaa H / Alkhaldi, Rami / Helal, Assala / Alamri, Sawsan S / Malki, Jana / Alhabbab, Rowa Y / Abujamel, Turki S / Alhakamy, Nabil A / Alnami, Aisha / Algaissi, Abdullah / Hassanain, Mazen / Hashem, Anwar M

Antibody therapeutics

2023 Volume 7, Issue 1, Page(s) 53–66

Abstract: The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and the Middle East respiratory syndrome coronavirus (MERS-CoV) are highly pathogenic human coronaviruses (CoVs). Anti-CoVs mAbs and vaccines may be effective, but the emergence of ... ...

Abstract	The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and the Middle East respiratory syndrome coronavirus (MERS-CoV) are highly pathogenic human coronaviruses (CoVs). Anti-CoVs mAbs and vaccines may be effective, but the emergence of neutralization escape variants is inevitable. Angiotensin-converting enzyme 2 and dipeptidyl peptidase 4 enzyme are the getaway receptors for SARS-CoV-2 and MERS-CoV, respectively. Thus, we reformatted these receptors as Fc-fusion decoy receptors. Then, we tested them in parallel with anti-SARS-CoV (ab1-IgG) and anti-MERS-CoV (M336-IgG) mAbs against several variants using pseudovirus neutralization assay. The generated Fc-based decoy receptors exhibited a strong inhibitory effect against all pseudotyped CoVs. Results showed that although mAbs can be effective antiviral drugs, they might rapidly lose their efficacy against highly mutated viruses. We suggest that receptor traps can be engineered as Fc-fusion proteins for highly mutating viruses with known entry receptors, for a faster and effective therapeutic response even against virus harboring antibodies escape mutations.
Language	English
Publishing date	2023-12-12
Publishing country	United States
Document type	Journal Article
ISSN	2516-4236
ISSN (online)	2516-4236
DOI	10.1093/abt/tbad030
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Article: Biosynthesis of 2-aminooctanoic acid and its use to terminally modify a lactoferricin B peptide derivative for improved antimicrobial activity

Almahboub, SarahA / Tanja Narancic / Marc Devocelle / Shane T. Kenny / William Palmer-Brown / Cormac Murphy / Jasmina Nikodinovic-Runic / Kevin E. O’Connor

Applied microbiology and biotechnology. 2018 Jan., v. 102, no. 2

2018

Abstract: Terminal modification of peptides is frequently used to improve their hydrophobicity. While N-terminal modification with fatty acids (lipidation) has been reported previously, C-terminal lipidation is limited as it requires the use of linkers. Here we ... ...

Abstract	Terminal modification of peptides is frequently used to improve their hydrophobicity. While N-terminal modification with fatty acids (lipidation) has been reported previously, C-terminal lipidation is limited as it requires the use of linkers. Here we report the use of a biocatalyst for the production of an unnatural fatty amino acid, (S)-2-aminooctanoic acid (2-AOA) with enantiomeric excess > 98% ee and the subsequent use of 2-AOA to modify and improve the activity of an antimicrobial peptide. A transaminase originating from Chromobacterium violaceum was employed with a conversion efficiency 52–80% depending on the ratio of amino group donor to acceptor. 2-AOA is a fatty acid with amino functionality, which allowed direct C- and N-terminal conjugation respectively to an antimicrobial peptide (AMP) derived from lactoferricin B. The antibacterial activity of the modified peptides was improved by up to 16-fold. Furthermore, minimal inhibitory concentrations (MIC) of C-terminally modified peptide were always lower than N-terminally conjugated peptides. The C-terminally modified peptide exhibited MIC values of 25 μg/ml for Escherichia coli, 50 μg/ml for Bacillus subtilis, 100 μg/ml for Salmonella typhimurium, 200 μg/ml for Pseudomonas aeruginosa and 400 μg/ml for Staphylococcus aureus. The C-terminally modified peptide was the only peptide tested that showed complete inhibition of growth of S. aureus.
Keywords	Bacillus subtilis ; Chromobacterium violaceum ; Escherichia coli ; Pseudomonas aeruginosa ; Salmonella Typhimurium ; Staphylococcus aureus ; amino acids ; antibacterial properties ; antimicrobial peptides ; biocatalysts ; biosynthesis ; fatty acids ; hydrophobicity ; lactoferrin ; minimum inhibitory concentration
Language	English
Dates of publication	2018-01
Size	p. 789-799.
Publishing place	Springer Berlin Heidelberg
Document type	Article
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-017-8655-0
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Seroprevalence of MERS-CoV in healthy adults in western Saudi Arabia, 2011-2016.

Degnah, Afnan A / Al-Amri, Sawsan S / Hassan, Ahmed M / Almasoud, Abdulrahman S / Mousa, Manar / Almahboub, Sarah A / Alhabbab, Rowa Y / Mirza, Ahmed A / Hindawi, Salwa I / Alharbi, Naif Khalaf / Azhar, Esam I / Hashem, Anwar M

Journal of infection and public health

2020 Volume 13, Issue 5, Page(s) 697–703

Abstract: Background: The Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly recognized zoonotic coronavirus. Current evidence confirms the role of dromedaries in primary human infections but does not explain the sporadic community cases. However, ...

Abstract	Background: The Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly recognized zoonotic coronavirus. Current evidence confirms the role of dromedaries in primary human infections but does not explain the sporadic community cases. However, asymptomatic or subclinical cases could represent a possible source of infection in the community. Methods: Archived human sera (7461) collected between 2011 and 2016 from healthy adult blood donors from 50 different nationalities in the western part of Saudi Arabia were obtained for MERS-CoV seroprevalence investigation. Samples were tested for MERS-CoV S1-specific antibodies (Abs) by ELISA and confirmed by testing for neutralizing Abs (nAbs) using both pseudotyped and live virus neutralization assays. Results: Out of 7461 samples, 174 sera from individuals with 18 different nationalities were ELISA positive (2.3%, 95% CI 2.0-2.7). Presence of nAbs was confirmed in 17 samples (0.23%, 95% CI 0.1-0.4) of which one sample exhibited positivity in both neutralization assays. Confirmed seropositivity was identified in young (15-44 years) men and women from Saudi Arabia, Egypt, Yemen, Pakistan, Palestine, Sudan, and India without significant preference. Conclusions: An increasing trend of MERS-CoV seroprevalence was observed in the general population in western Saudi Arabia, suggesting that asymptomatic or mild infections might exist and act as an unrecognized source of infection. Seropositivity of individuals from different nationalities underscores the potential MERS exportation outside of the Arabian Peninsula. Thus, enhanced and continuous surveillance is highly warranted.
MeSH term(s)	Adolescent ; Adult ; Animals ; Antibodies, Viral/blood ; Blood Donors ; Camelus/virology ; Coronavirus Infections/blood ; Coronavirus Infections/epidemiology ; Coronavirus Infections/immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Middle East Respiratory Syndrome Coronavirus/immunology ; Middle East Respiratory Syndrome Coronavirus/isolation & purification ; Saudi Arabia/epidemiology ; Seroepidemiologic Studies ; Young Adult
Chemical Substances	Antibodies, Viral
Keywords	covid19
Language	English
Publishing date	2020-01-28
Publishing country	England
Document type	Journal Article
ISSN	1876-035X
ISSN (online)	1876-035X
DOI	10.1016/j.jiph.2020.01.001
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Seroprevalence of MERS-CoV in healthy adults in western Saudi Arabia, 2011-2016

J Infect Public Health

Abstract: BACKGROUND: The Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly recognized zoonotic coronavirus. Current evidence confirms the role of dromedaries in primary human infections but does not explain the sporadic community cases. However, ... ...

Abstract	BACKGROUND: The Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly recognized zoonotic coronavirus. Current evidence confirms the role of dromedaries in primary human infections but does not explain the sporadic community cases. However, asymptomatic or subclinical cases could represent a possible source of infection in the community. METHODS: Archived human sera (7461) collected between 2011 and 2016 from healthy adult blood donors from 50 different nationalities in the western part of Saudi Arabia were obtained for MERS-CoV seroprevalence investigation. Samples were tested for MERS-CoV S1-specific antibodies (Abs) by ELISA and confirmed by testing for neutralizing Abs (nAbs) using both pseudotyped and live virus neutralization assays. RESULTS: Out of 7461 samples, 174 sera from individuals with 18 different nationalities were ELISA positive (2.3%, 95% CI 2.0-2.7). Presence of nAbs was confirmed in 17 samples (0.23%, 95% CI 0.1-0.4) of which one sample exhibited positivity in both neutralization assays. Confirmed seropositivity was identified in young (15-44 years) men and women from Saudi Arabia, Egypt, Yemen, Pakistan, Palestine, Sudan, and India without significant preference. CONCLUSIONS: An increasing trend of MERS-CoV seroprevalence was observed in the general population in western Saudi Arabia, suggesting that asymptomatic or mild infections might exist and act as an unrecognized source of infection. Seropositivity of individuals from different nationalities underscores the potential MERS exportation outside of the Arabian Peninsula. Thus, enhanced and continuous surveillance is highly warranted.
Keywords	covid19
Publisher	WHO
Document type	Article
Note	WHO #Covidence: #154833
Database	COVID19

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Article ; Online: Biosynthesis of 2-aminooctanoic acid and its use to terminally modify a lactoferricin B peptide derivative for improved antimicrobial activity.

Almahboub, Sarah A / Narancic, Tanja / Devocelle, Marc / Kenny, Shane T / Palmer-Brown, William / Murphy, Cormac / Nikodinovic-Runic, Jasmina / O'Connor, Kevin E

Applied microbiology and biotechnology

2017 Volume 102, Issue 2, Page(s) 789–799

Abstract	Terminal modification of peptides is frequently used to improve their hydrophobicity. While N-terminal modification with fatty acids (lipidation) has been reported previously, C-terminal lipidation is limited as it requires the use of linkers. Here we report the use of a biocatalyst for the production of an unnatural fatty amino acid, (S)-2-aminooctanoic acid (2-AOA) with enantiomeric excess > 98% ee and the subsequent use of 2-AOA to modify and improve the activity of an antimicrobial peptide. A transaminase originating from Chromobacterium violaceum was employed with a conversion efficiency 52-80% depending on the ratio of amino group donor to acceptor. 2-AOA is a fatty acid with amino functionality, which allowed direct C- and N-terminal conjugation respectively to an antimicrobial peptide (AMP) derived from lactoferricin B. The antibacterial activity of the modified peptides was improved by up to 16-fold. Furthermore, minimal inhibitory concentrations (MIC) of C-terminally modified peptide were always lower than N-terminally conjugated peptides. The C-terminally modified peptide exhibited MIC values of 25 μg/ml for Escherichia coli, 50 μg/ml for Bacillus subtilis, 100 μg/ml for Salmonella typhimurium, 200 μg/ml for Pseudomonas aeruginosa and 400 μg/ml for Staphylococcus aureus. The C-terminally modified peptide was the only peptide tested that showed complete inhibition of growth of S. aureus.
MeSH term(s)	Alkynes/chemistry ; Amino Acids/biosynthesis ; Anti-Infective Agents/pharmacology ; Antimicrobial Cationic Peptides/chemistry ; Antimicrobial Cationic Peptides/pharmacology ; Bacillus subtilis/drug effects ; Biocatalysis ; Caprylates/chemistry ; Chromobacterium/enzymology ; Hydrophobic and Hydrophilic Interactions ; Lactoferrin/chemistry ; Microbial Sensitivity Tests ; Pseudomonas aeruginosa/drug effects ; Staphylococcus aureus/drug effects ; Transaminases/metabolism
Chemical Substances	2-aminooctynoic acid ; Alkynes ; Amino Acids ; Anti-Infective Agents ; Antimicrobial Cationic Peptides ; Caprylates ; lactoferricin B (146897-68-9) ; Transaminases (EC 2.6.1.-) ; Lactoferrin (EC 3.4.21.-)
Language	English
Publishing date	2017-11-25
Publishing country	Germany
Document type	Journal Article
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-017-8655-0
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