Quantitative real-time PCR monitoring dynamics of Thermotoga neapolitana in synthetic co-culture for biohydrogen production

Onyinye Okonkwo; Aino Maija Lakaniemi; Ville Santala; Matti Karp; Rahul Mangayil

doi:10.1016/j.ijhydene.2017.12.002

Quantitative real-time PCR monitoring dynamics of Thermotoga neapolitana in synthetic co-culture for biohydrogen production

Onyinye Okonkwo^*
, Aino Maija Lakaniemi
, Ville Santala
, Matti Karp
, Rahul Mangayil

^*Corresponding author for this work

Not published at Aalto University

Tampere University

Research output: Contribution to journal › Article › Scientific › peer-review

18 Citations (Scopus)

Abstract

This study demonstrates the potential for biohydrogen production in a co-culture of two ecologically distant species, Thermatoga neapolitana and Caldicellulosiruptor saccharolyticus, and the development of a quantitative real-time PCR (qPCR) method for quantifying the hyperthermophilic bacterium of the genus Thermotoga. Substrate utilization and H₂ production performance was compared to those of their individual cultures. The highest H₂ yields obtained were 2.7 ± 0.05, 2.5 ± 0.07 and 2.8 ± 0.09 mol H₂/mol glucose for C. saccharolyticus, T. neapolitana, and their co-culture respectively. Statistical analysis comparing the H₂ production rate of the co-culture to either C. saccahrolyticus or T. neapolitana pure cultures indicated a significant difference in the H₂ production rate (p < 0.05: t-test), with the highest rate of H₂ production (36.02 mL L⁻¹ h⁻¹) observed from the co-culture fermentations. In order to monitor the presence of T. neapolitana in the bioprocess, we developed a qPCR method using 16S rRNA gene and hydrogenase (hydA) gene targets. The qPCR data using hydA primers specific to T. neapolitana showed an increase in hydA gene copies from 3.32 × 10⁷ to 4.4 × 10⁸ hydA gene copies per mL confirming the influence of T. neapolitana in the synthetic consortium.

Original language	English
Pages (from-to)	3133-3141
Number of pages	9
Journal	International Journal of Hydrogen Energy
Volume	43
Issue number	6
DOIs	https://doi.org/10.1016/j.ijhydene.2017.12.002
Publication status	Published - 8 Feb 2018
MoE publication type	A1 Journal article-refereed

Funding

This work was supported by the Marie Skłodowska-Curie European Joint Doctorate (EJD) in Advanced Biological Waste-To-Energy Technologies (ABWET) funded from Horizon 2020 under grant agreement no. 643071 . This work was supported by the Marie Skłodowska-Curie European Joint Doctorate (EJD) in Advanced Biological Waste-To-Energy Technologies (ABWET) funded from Horizon 2020 under grant agreement no. 643071.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 7 Affordable and Clean Energy

Keywords

16S rRNA gene
hydA
Hydrogen production
Primer design
Real-time quantitative polymerase chain reaction
Thermotoga neapolitana

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10.1016/j.ijhydene.2017.12.002

Cite this

@article{54d1495a2c814a1ca92f74820bacd85e,

title = "Quantitative real-time PCR monitoring dynamics of Thermotoga neapolitana in synthetic co-culture for biohydrogen production",

abstract = "This study demonstrates the potential for biohydrogen production in a co-culture of two ecologically distant species, Thermatoga neapolitana and Caldicellulosiruptor saccharolyticus, and the development of a quantitative real-time PCR (qPCR) method for quantifying the hyperthermophilic bacterium of the genus Thermotoga. Substrate utilization and H2 production performance was compared to those of their individual cultures. The highest H2 yields obtained were 2.7 ± 0.05, 2.5 ± 0.07 and 2.8 ± 0.09 mol H2/mol glucose for C. saccharolyticus, T. neapolitana, and their co-culture respectively. Statistical analysis comparing the H2 production rate of the co-culture to either C. saccahrolyticus or T. neapolitana pure cultures indicated a significant difference in the H2 production rate (p < 0.05: t-test), with the highest rate of H2 production (36.02 mL L−1 h−1) observed from the co-culture fermentations. In order to monitor the presence of T. neapolitana in the bioprocess, we developed a qPCR method using 16S rRNA gene and hydrogenase (hydA) gene targets. The qPCR data using hydA primers specific to T. neapolitana showed an increase in hydA gene copies from 3.32 × 107 to 4.4 × 108 hydA gene copies per mL confirming the influence of T. neapolitana in the synthetic consortium.",

keywords = "16S rRNA gene, hydA, Hydrogen production, Primer design, Real-time quantitative polymerase chain reaction, Thermotoga neapolitana",

author = "Onyinye Okonkwo and Lakaniemi, \{Aino Maija\} and Ville Santala and Matti Karp and Rahul Mangayil",

note = "Funding Information: This work was supported by the Marie Sk{\l}odowska-Curie European Joint Doctorate (EJD) in Advanced Biological Waste-To-Energy Technologies (ABWET) funded from Horizon 2020 under grant agreement no. 643071 . Funding Information: This work was supported by the Marie Sk{\l}odowska-Curie European Joint Doctorate (EJD) in Advanced Biological Waste-To-Energy Technologies (ABWET) funded from Horizon 2020 under grant agreement no. 643071. Publisher Copyright: {\textcopyright} 2017 Hydrogen Energy Publications LLC",

year = "2018",

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doi = "10.1016/j.ijhydene.2017.12.002",

language = "English",

volume = "43",

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journal = "International Journal of Hydrogen Energy",

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T1 - Quantitative real-time PCR monitoring dynamics of Thermotoga neapolitana in synthetic co-culture for biohydrogen production

AU - Okonkwo, Onyinye

AU - Lakaniemi, Aino Maija

AU - Santala, Ville

AU - Karp, Matti

AU - Mangayil, Rahul

N1 - Funding Information: This work was supported by the Marie Skłodowska-Curie European Joint Doctorate (EJD) in Advanced Biological Waste-To-Energy Technologies (ABWET) funded from Horizon 2020 under grant agreement no. 643071 . Funding Information: This work was supported by the Marie Skłodowska-Curie European Joint Doctorate (EJD) in Advanced Biological Waste-To-Energy Technologies (ABWET) funded from Horizon 2020 under grant agreement no. 643071. Publisher Copyright: © 2017 Hydrogen Energy Publications LLC

PY - 2018/2/8

Y1 - 2018/2/8

N2 - This study demonstrates the potential for biohydrogen production in a co-culture of two ecologically distant species, Thermatoga neapolitana and Caldicellulosiruptor saccharolyticus, and the development of a quantitative real-time PCR (qPCR) method for quantifying the hyperthermophilic bacterium of the genus Thermotoga. Substrate utilization and H2 production performance was compared to those of their individual cultures. The highest H2 yields obtained were 2.7 ± 0.05, 2.5 ± 0.07 and 2.8 ± 0.09 mol H2/mol glucose for C. saccharolyticus, T. neapolitana, and their co-culture respectively. Statistical analysis comparing the H2 production rate of the co-culture to either C. saccahrolyticus or T. neapolitana pure cultures indicated a significant difference in the H2 production rate (p < 0.05: t-test), with the highest rate of H2 production (36.02 mL L−1 h−1) observed from the co-culture fermentations. In order to monitor the presence of T. neapolitana in the bioprocess, we developed a qPCR method using 16S rRNA gene and hydrogenase (hydA) gene targets. The qPCR data using hydA primers specific to T. neapolitana showed an increase in hydA gene copies from 3.32 × 107 to 4.4 × 108 hydA gene copies per mL confirming the influence of T. neapolitana in the synthetic consortium.

AB - This study demonstrates the potential for biohydrogen production in a co-culture of two ecologically distant species, Thermatoga neapolitana and Caldicellulosiruptor saccharolyticus, and the development of a quantitative real-time PCR (qPCR) method for quantifying the hyperthermophilic bacterium of the genus Thermotoga. Substrate utilization and H2 production performance was compared to those of their individual cultures. The highest H2 yields obtained were 2.7 ± 0.05, 2.5 ± 0.07 and 2.8 ± 0.09 mol H2/mol glucose for C. saccharolyticus, T. neapolitana, and their co-culture respectively. Statistical analysis comparing the H2 production rate of the co-culture to either C. saccahrolyticus or T. neapolitana pure cultures indicated a significant difference in the H2 production rate (p < 0.05: t-test), with the highest rate of H2 production (36.02 mL L−1 h−1) observed from the co-culture fermentations. In order to monitor the presence of T. neapolitana in the bioprocess, we developed a qPCR method using 16S rRNA gene and hydrogenase (hydA) gene targets. The qPCR data using hydA primers specific to T. neapolitana showed an increase in hydA gene copies from 3.32 × 107 to 4.4 × 108 hydA gene copies per mL confirming the influence of T. neapolitana in the synthetic consortium.

KW - 16S rRNA gene

KW - hydA

KW - Hydrogen production

KW - Primer design

KW - Real-time quantitative polymerase chain reaction

KW - Thermotoga neapolitana

UR - https://www.scopus.com/pages/publications/85041384509

U2 - 10.1016/j.ijhydene.2017.12.002

DO - 10.1016/j.ijhydene.2017.12.002

M3 - Article

AN - SCOPUS:85041384509

SN - 0360-3199

VL - 43

SP - 3133

EP - 3141

JO - International Journal of Hydrogen Energy

JF - International Journal of Hydrogen Energy

IS - 6

ER -

Quantitative real-time PCR monitoring dynamics of Thermotoga neapolitana in synthetic co-culture for biohydrogen production

Abstract

Funding

UN SDGs

Keywords

Access to Document

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