Abstract
Parachlorella kessleri (formerly Chlorella kessleri) can accumulate high levels of both starch and lipids that can be used for the production of bioethanol and biodiesel, respectively. We mutagenized P. kessleri by heavy-ion beam irradiation and obtained a biotechnologically promising mutant strain, PK4, for further improvement of lipid productivity in comparison with wild type [1]. PK4 reached two times the optical density and accumulated 2.6 times as much starch in complete urea-phosphate (UP) media compared to tris-acetate-phosphate (TAP) media under laboratory conditions. Similarly to wild type, PK4 accumulated only negligible amounts of lipids in complete UP media. PK4 accumulated more lipids (WT: 1.17 g L−1, PK4: 1.75 g L−1) and accumulated lipids faster than WT after dilution of the UP media (WT; 0.22 g L−1 day−1, PK4; 0.43 g L−1 day−1). The productivity of PK4 was analyzed in mass culture using a 150-L thin-layer photo bioreactor housed in a temperature and light-uncontrolled glass greenhouse. The PBR consisted of two glass plates (each 6 m long and 1 m wide) at an inclination of 1.6%, arranged in a meandering path and connected with a trough. The culture was diluted four-fold with water on day 7 post-inoculation to ensure prompt nutrient limitation in the UP media. Lipid accumulation in PK4 was significantly induced so that it accumulated 66% of lipid per dry weight. Under 150-L mass cultivation conditions, PK4 showed high biomass productivity (0.82 g L−1 day−1) and high lipid productivity (0.59 g L−1 day−1). In addition, the whole genome of PK4 was sequenced to clarify the genetic variation upon heavy-ion-beam irradiation by comparison with the reference genome of WT. As a result, genetic differentiation of PK4 was found at three genes encoding endo-1,4-β-mannanase, an ATP/ADP transporter, and an elicitor response protein.
| Original language | English |
|---|---|
| Pages (from-to) | 416-426 |
| Number of pages | 11 |
| Journal | Algal Research |
| Volume | 35 |
| DOIs | |
| Publication status | Published - 2018 Nov 1 |
| Externally published | Yes |
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Keywords
- 150-L mass cultivation
- Mutation identification
- Nutrient dilution
- Parachlorella kessleri
- Single-nucleotide polymorphism
- Whole genome sequencing
ASJC Scopus subject areas
- Agronomy and Crop Science
Cite this
Comparison of lipid productivity of Parachlorella kessleri heavy-ion beam irradiation mutant PK4 in laboratory and 150-L mass bioreactor, identification and characterization of its genetic variation. / Takeshita, Tsuyoshi; Ivanov, Ivan Nedyalkov; Oshima, Kenshiro; Ishii, Kotaro; Kawamoto, Hiroki; Ota, Shuhei; Yamazaki, Tomokazu; Hirata, Aiko; Kazama, Yusuke; Abe, Tomoko; Hattori, Masahira; Bišová, Kateřina; Zachleder, Vilém; Kawano, Shigeyuki.
In: Algal Research, Vol. 35, 01.11.2018, p. 416-426.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Comparison of lipid productivity of Parachlorella kessleri heavy-ion beam irradiation mutant PK4 in laboratory and 150-L mass bioreactor, identification and characterization of its genetic variation
AU - Takeshita, Tsuyoshi
AU - Ivanov, Ivan Nedyalkov
AU - Oshima, Kenshiro
AU - Ishii, Kotaro
AU - Kawamoto, Hiroki
AU - Ota, Shuhei
AU - Yamazaki, Tomokazu
AU - Hirata, Aiko
AU - Kazama, Yusuke
AU - Abe, Tomoko
AU - Hattori, Masahira
AU - Bišová, Kateřina
AU - Zachleder, Vilém
AU - Kawano, Shigeyuki
PY - 2018/11/1
Y1 - 2018/11/1
N2 - Parachlorella kessleri (formerly Chlorella kessleri) can accumulate high levels of both starch and lipids that can be used for the production of bioethanol and biodiesel, respectively. We mutagenized P. kessleri by heavy-ion beam irradiation and obtained a biotechnologically promising mutant strain, PK4, for further improvement of lipid productivity in comparison with wild type [1]. PK4 reached two times the optical density and accumulated 2.6 times as much starch in complete urea-phosphate (UP) media compared to tris-acetate-phosphate (TAP) media under laboratory conditions. Similarly to wild type, PK4 accumulated only negligible amounts of lipids in complete UP media. PK4 accumulated more lipids (WT: 1.17 g L−1, PK4: 1.75 g L−1) and accumulated lipids faster than WT after dilution of the UP media (WT; 0.22 g L−1 day−1, PK4; 0.43 g L−1 day−1). The productivity of PK4 was analyzed in mass culture using a 150-L thin-layer photo bioreactor housed in a temperature and light-uncontrolled glass greenhouse. The PBR consisted of two glass plates (each 6 m long and 1 m wide) at an inclination of 1.6%, arranged in a meandering path and connected with a trough. The culture was diluted four-fold with water on day 7 post-inoculation to ensure prompt nutrient limitation in the UP media. Lipid accumulation in PK4 was significantly induced so that it accumulated 66% of lipid per dry weight. Under 150-L mass cultivation conditions, PK4 showed high biomass productivity (0.82 g L−1 day−1) and high lipid productivity (0.59 g L−1 day−1). In addition, the whole genome of PK4 was sequenced to clarify the genetic variation upon heavy-ion-beam irradiation by comparison with the reference genome of WT. As a result, genetic differentiation of PK4 was found at three genes encoding endo-1,4-β-mannanase, an ATP/ADP transporter, and an elicitor response protein.
AB - Parachlorella kessleri (formerly Chlorella kessleri) can accumulate high levels of both starch and lipids that can be used for the production of bioethanol and biodiesel, respectively. We mutagenized P. kessleri by heavy-ion beam irradiation and obtained a biotechnologically promising mutant strain, PK4, for further improvement of lipid productivity in comparison with wild type [1]. PK4 reached two times the optical density and accumulated 2.6 times as much starch in complete urea-phosphate (UP) media compared to tris-acetate-phosphate (TAP) media under laboratory conditions. Similarly to wild type, PK4 accumulated only negligible amounts of lipids in complete UP media. PK4 accumulated more lipids (WT: 1.17 g L−1, PK4: 1.75 g L−1) and accumulated lipids faster than WT after dilution of the UP media (WT; 0.22 g L−1 day−1, PK4; 0.43 g L−1 day−1). The productivity of PK4 was analyzed in mass culture using a 150-L thin-layer photo bioreactor housed in a temperature and light-uncontrolled glass greenhouse. The PBR consisted of two glass plates (each 6 m long and 1 m wide) at an inclination of 1.6%, arranged in a meandering path and connected with a trough. The culture was diluted four-fold with water on day 7 post-inoculation to ensure prompt nutrient limitation in the UP media. Lipid accumulation in PK4 was significantly induced so that it accumulated 66% of lipid per dry weight. Under 150-L mass cultivation conditions, PK4 showed high biomass productivity (0.82 g L−1 day−1) and high lipid productivity (0.59 g L−1 day−1). In addition, the whole genome of PK4 was sequenced to clarify the genetic variation upon heavy-ion-beam irradiation by comparison with the reference genome of WT. As a result, genetic differentiation of PK4 was found at three genes encoding endo-1,4-β-mannanase, an ATP/ADP transporter, and an elicitor response protein.
KW - 150-L mass cultivation
KW - Mutation identification
KW - Nutrient dilution
KW - Parachlorella kessleri
KW - Single-nucleotide polymorphism
KW - Whole genome sequencing
UR - http://www.scopus.com/inward/record.url?scp=85053840822&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85053840822&partnerID=8YFLogxK
U2 - 10.1016/j.algal.2018.09.005
DO - 10.1016/j.algal.2018.09.005
M3 - Article
VL - 35
SP - 416
EP - 426
JO - Algal Research
JF - Algal Research
SN - 2211-9264
ER -