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The Transport Behavior of a Biflagellated Microswimmer before and after Cargo Loading

Teng XJ, Ng WM, Chong WH, Chan DJC, Mohamud R, Ooi BS, et al.

Langmuir, 2021 08 03;37(30):9192-9201.
PMID: 34255525 DOI: 10.1021/acs.langmuir.1c01345

The changes in the transport behavior of a microswimmer before and after cargo loading are crucial to understanding and control of the motion of a biohybrid microbot. In this work, we show the change in swimming behavior of biflagellated microalgae Chlamydomonas reinhardtii picking up a 4.5 μm polystyrene microbead upon collision. The microswimmer changed from linear forward motion into helical motion upon the attachment of the cargo and swam with a decreased swimming velocity. We revealed the helical motion of the microswimmer upon cargo loading due to suppression of flagella by image analysis of magnified time-lapse images of C. reinhardtii with one microbead attached at the anterior end (between the flagella). Furthered suppression on the flagellum imposed by the loading of the second cargo has led to increased oscillation per displacement traveled and decreased swimming velocity. Moreover, the microswimmer with a microbead attached at the posterior end swam with swimming velocity close to free swimming microalgae and did not exhibit helical swimming behavior. The experimental results and analysis showed that the loading location of the cargo has a great influence over the swimming behavior of the microswimmer. Furthermore, the work balance calculation and mathematical analysis based on Lighthill's model are well consistent with our experimental findings.

Matched MeSH terms: Chlamydomonas reinhardtii*
Dissection of synechococcus rubisco large subunit sections involved in holoenzyme formation in escherichia coli by combinatorial section swapping and sequence analyses

Yee Hung Yeap, Teng Wei Koay, Boon Hoe Lim

Sains Malaysiana, 2018;47:2269-2289.

Engineering the CO2
-fixing enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) to improve photosynthesis
has long been sought. Rubisco large subunits (RbcL) are highly-conserved but because of certain undefined sequence
differences, plant Rubisco research cannot fully utilise the robust heterologous Escherichia coli expression system and its
GroEL folding machinery. Previously, a series of chimeric cyanobacteria Synechococcus elongatus Rubisco, incorporated
with sequences from the green alga Chlamydomonas reinhardtii, were expressed in E. coli; differences in RbcL sections
essential for holoenzyme formation were pinpointed. In this study, the remaining sections, presumably not crucial for
holoenzyme formation and also the small subunit (RbcS), are substituted to further ascertain the possible destabilising
effects of multiple section mutations. To that end, combinations of Synechococcus RbcL Sections 1 (residues 1-47), 2
(residues 48-97), 5 (residues 198-247) and 10 (residues 448-472), and RbcS, were swapped with collinear Chlamydomonas
sections and expressed in E. coli. Interestingly, only the chimera with Sections 1 and 2 together produces holoenzyme and
an interaction network of complementing amino acid changes is delineated by crystal structure analysis. Furthermore,
sequence-based analysis also highlighted possible GroEL binding site differences between the two RbcLs.

Matched MeSH terms: Chlamydomonas; Chlamydomonas reinhardtii
Biosorption of copper and zinc by immobilised and free algal biomass, and the effects of metal biosorption on the growth and cellular structure of Chlorella sp. and Chlamydomonas sp. isolated from rivers in Penang, Malaysia

Maznah WO, Al-Fawwaz AT, Surif M

J Environ Sci (China), 2012;24(8):1386-93.
PMID: 23513679

In this study, the biosorption of copper and zinc ions by Chlorella sp. and Chlamydomonas sp. isolated from local environments in Malaysia was investigated in a batch system and by microscopic analyses. Under optimal biosorption conditions, the biosorption capacity of Chlorella sp. for copper and zinc ions was 33.4 and 28.5 mg/g, respectively, after 6 hr of biosorption in an immobilised system. Batch experiments showed that the biosorption capacity of algal biomass immobilised in the form of sodium alginate beads was higher than that of the free biomass. Scanning electron microscopy and energy-dispersive X-ray spectroscopy analyses revealed that copper and zinc were mainly sorbed at the cell surface during biosorption. Exposure to 5 mg/L of copper and zinc affected both the chlorophyll content and cell count of the algal cells after the first 12 hr of contact time.

Matched MeSH terms: Chlamydomonas/drug effects; Chlamydomonas/growth & development; Chlamydomonas/chemistry*
Fulltext Marine microalgae co-cultured with floc-forming bacterium: Insight into growth and lipid productivity

Yee CS, Okomoda VT, Hashim F, Waiho K, Sheikh Abdullah SR, Alamanjo C, et al.

PeerJ, 2021;9:e11217.
PMID: 33981498 DOI: 10.7717/peerj.11217

This study investigated the effect of co-culturing microalgae with a floc-forming bacterium. Of the six microalgae isolated from a biofloc sample, only Thalassiosira weissflogii, Chlamydomonas sp. and Chlorella vulgaris were propagated successfully in Conway medium. Hence, these species were selected for the experiment comparing microalgae axenic culture and co-culture with the floc-forming bacterium, Bacillus infantis. Results obtained showed that the co-culture had higher microalgae biomass compared to the axenic culture. A similar trend was also observed concerning the lipid content of the microalgae-bacterium co-cultures. The cell number of B. infantis co-cultured with T. weissflogii increased during the exponential stage until the sixth day, but the other microalgae species experienced a significant early reduction in cell density of the bacteria at the exponential stage. This study represents the first attempt at co-culturing microalgae with B. infantis, a floc-forming bacterium, and observed increased biomass growth and lipid accumulation compared to the axenic culture.

Matched MeSH terms: Chlamydomonas
Exploring the potency of integrating semi-batch operation into lipid yield performance of Chlamydomonas sp. Tai-03

Tan CH, Show PL, Ling TC, Nagarajan D, Lee DJ, Chen WH, et al.

Bioresour Technol, 2019 Aug;285:121331.
PMID: 30999192 DOI: 10.1016/j.biortech.2019.121331

Third generation biofuels, also known as microalgal biofuels, are promising alternatives to fossil fuels. One attractive option is microalgal biodiesel as a replacement for diesel fuel. Chlamydomonas sp. Tai-03 was previously optimized for maximal lipid production for biodiesel generation, achieving biomass growth and productivity of 3.48 ± 0.04 g/L and 0.43 ± 0.01 g/L/d, with lipid content and productivity of 28.6 ± 1.41% and 124.1 ± 7.57 mg/L/d. In this study, further optimization using 5% CO2 concentration and semi-batch operation with 25% medium replacement ratio, enhanced the biomass growth and productivity to 4.15 ± 0.12 g/L and 1.23 ± 0.02 g/L/d, with lipid content and productivity of 19.4 ± 2.0% and 239.6 ± 24.8 mg/L/d. The major fatty acid methyl esters (FAMEs) were palmitic acid (C16:0), oleic acid (C18:1), and linoleic acid (C18:2). These short-chain FAMEs combined with high growth make Chlamydomonas sp. Tai-03 a suitable candidate for biodiesel synthesis.

Matched MeSH terms: Chlamydomonas
Fulltext A simple and inexpensive physical lysis method for DNA and RNA extraction from freshwater microalgae

Yee W, Abdul-Kadir R, Lee LM, Koh B, Lee YS, Chan HY

3 Biotech, 2018 Aug;8(8):354.
PMID: 30105179 DOI: 10.1007/s13205-018-1381-1

In this work, a simple and inexpensive physical lysis method using a cordless drill fitted with a plastic pellet pestle and 150 mg of sterile sea sand was established for the extraction of DNA from six strains of freshwater microalgae. This lysis method was also tested for RNA extraction from two microalgal strains. Lysis duration between 15 and 120 s using the cetyltrimethyl ammonium bromide (CTAB) buffer significantly increased the yield of DNA from four microalgalstrains (Monoraphidium griffithii NS16, Scenedesmus sp. NS6, Scenedesmus sp. DPBC1 and Acutodesmus sp. DPBB10) compared to control. It was also found that grinding was not required to obtain DNA from two strains of microalgae (Choricystis sp. NPA14 and Chlamydomonas sp. BM3). The average DNA yield obtained using this lysis method was between 62.5 and 78.9 ng/mg for M. griffithii NS16, 42.2-247.0 ng/mg for Scenedesmus sp. NS6, 70.2-110.9 ng/mg for Scenedesmus sp. DPBC1 and 142.8-164.8 ng/mg for Acutodesmus sp. DPBB10. DNA obtained using this method was sufficiently pure for PCR amplification. Extraction of total RNA from M. griffithii NS16 and Mychonastes sp. NPD7 using this lysis method yielded high-quality RNA suitable for RT-PCR. This lysis method is simple, cheap and would enable rapid nucleic acid extraction from freshwater microalgae without requiring costly materials and equipment such as liquid nitrogen or beadbeaters, and would facilitate molecular studies on microalgae in general.

Matched MeSH terms: Chlamydomonas
Integrated palm oil mill effluent treatment and co2 sequestration by microalgae

Nur Anira Syafiqah Hazman, Hassimi Abu Hasan, Kamrul Fakir Kamarudin, Nazlina Haiza Mohd Yasin, Mohd Sobri Takriff, Noor Irma Nazashida Mohd Hakimi

Sains Malaysiana, 2018;47:1455-1464.

Malaysian economy relies on palm oil industries as a driver for rural development. However, palm oil mill effluent
(POME) that is generated from palm oil processing stages causes major environmental challenges. Before being
released to the environment, POME treatment is crucial to comply with standard discharge limit. Microalgae have
demonstrated excellent potential for phycoremediating POME and capturing CO2
. In this study, local microalgae isolate
such as Chlamydomonas sp. UKM 6 and Chlorella spp. UKM 8 were used for POME treatment in 21 days with different
inoculum sizes (5%, 10% and 15%). In addition, an integrated treatment process was performed by taking the treated
POME supernatant for cultivation of Chorella spp. UKM 2, Chorella sorokiniana UKM 3 and Chlorella vulgaris for CO2
sequestration study. Different CO2
concentrations (5%, 10% and 15%) were used and the experiments were carried
out in 10 days under continuous illumination. The results showed that among two species involves in POME treatment,
Chlamydomonas sp. UKM 6 showed a great potential to remove pollutant such as COD (56%), nitrogen (65%) and
phosphorus (34%). The biomass after POME treatment and CO2
biofixation content high lipid (90 mg lipid/g biomass)
which can be the potential source for biodiesel production. In CO2
sequestration study, C. sorokininana UKM3 able
to takes up to 15% CO2
with CO2
uptake rate of 273 mgL-1d-1. In this study, the integrated system of POME treatment
and CO2
sequestration were feasible using microalgae.

Matched MeSH terms: Chlamydomonas
Characterization of Chlamydomonas Ribulose-1,5-bisphosphate carboxylase/oxygenase variants mutated at residues that are post-translationally modified

Rasineni GK, Loh PC, Lim BH

Biochim Biophys Acta Gen Subj, 2017 Feb;1861(2):79-85.
PMID: 27816753 DOI: 10.1016/j.bbagen.2016.10.027

BACKGROUND: Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is the chloroplast enzyme that fixes CO2 in photosynthesis, but the enzyme also fixes O2, which leads to the wasteful photorespiratory pathway. If we better understand the structure-function relationship of the enzyme, we might be able to engineer improvements. When the crystal structure of Chlamydomonas Rubisco was solved, four new posttranslational modifications were observed which are not present in other species. The modifications were 4-hydroxylation of the conserved Pro-104 and 151 residues, and S-methylation of the variable Cys-256 and 369 residues, which are Phe-256 and Val-369 in land plants. Because the modifications were only observed in Chlamydomonas Rubisco, they might account for the differences in kinetic properties between the algal and plant enzymes.
METHODS: Site-directed mutagenesis and chloroplast transformation have been used to test the essentiality of these modifications by replacing each of the residues with alanine (Ala). Biochemical analyses were done to determine the specificity factors and kinetic constants.
RESULTS: Replacing the modified-residues in Chlamydomonas Rubisco affected the enzyme's catalytic activity. Substituting hydroxy-Pro-104 and methyl-Cys-256 with alanine influenced Rubisco catalysis.
CONCLUSION: This is the first study on these posttranslationally-modified residues in Rubisco by genetic engineering. As these forms of modifications/regulation are not available in plants, the modified residues could be a means to modulate Rubisco activity.
GENERAL SIGNIFICANCE: With a better understanding of Rubisco structure-function, we can define targets for improving the enzyme.

Matched MeSH terms: Chlamydomonas reinhardtii/genetics*