Chun-Zhao Liu

Microalgal bioreactors: challenges and opportunities.

Cultivating and harvesting of products from microalgae has led to increasing commercial interest in their use for producing valuable substances for food, feed, cosmetics, pharmaceuticals, and biodiesel, as well as for mitigation of pollution and rising CO2 in the environment. This review outlines different bioreactors and their current status, and points out their advantages and disadvantages. Compared with open‐air systems, there are distinct advantages to using closed systems, but technical challenges still remain. In view of potential applications, development of a more controllable, economical, and efficient closed culturing system is needed. Further developments still depend on continued research in the design of photobioreactors and break‐throughs in microalgal culturing technologies.

A simple and rapid harvesting method for microalgae by in situ magnetic separation.

A simple and rapid harvesting method by in situ magnetic separation with naked Fe(3)O(4) nanoparticles has been developed for the microalgal recovery of Botryococcus braunii and Chlorella ellipsoidea. After adding the magnetic particles to the microalgal culture broth, the microalgal cells were adsorbed and then separated by an external magnetic field. The maximal recovery efficiency reached more than 98% for both microalgae at a stirring speed of 120 r/min within 1 min, and the maximal adsorption capacity of these Fe(3)O(4) nanoparticles reached 55.9 mg-dry biomass/mg-particles for B. braunii and 5.83 mg-dry biomass/mg-particles for C. ellipsoidea. Appropriate pH value and high nanoparticle dose were favorable to the microalgae recovery, and the adsorption mechanism between the naked Fe(3)O(4) nanoparticles and the microalgal cells was mainly due to the electrostatic attraction. The developed in situ magnetic separation technology provides a great potential for saving time and energy associated with improving microalgal harvesting.

Magnetic mesoporous silica nanoparticles: Fabrication and their laccase immobilization performance

Newly large-pore magnetic mesoporous silica nanoparticles (MMSNPs) with wormhole framework structures were synthesized for the first time by using tetraethyl orthosilicate as the silica source and amine-terminated Jeffamine surfactants as template. Iminodiacerate was attached on these MMSNPs through a silane-coupling agent and chelated with Cu(2+). The Cu(2+)-chelated MMSNPs (MMSNPs-CPTS-IDA-Cu(2+)) showed higher adsorption capacity of 98.1 mg g(-1)-particles and activity recovery of 92.5% for laccase via metal affinity adsorption in comparison with MMSNPs via physical adsorption. The Michaelis constant (K(m)) and catalytic constant (k(cat)) of laccase immobilized on the MMSNPs-CPTS-IDA-Cu(2+) were 3.28 mM and 155.4 min(-1), respectively. Storage stability and temperature endurance of the immobilized laccase on MMSNPs-CPTS-IDA-Cu(2+) increased significantly, and the immobilized laccase retained 86.6% of its initial activity after 10 successive batch reactions operated with magnetic separation.

Separation of chlorogenic acid from honeysuckle crude extracts by macroporous resins.

Chlorogenic acid, one of the most bioactive compounds rich in the Chinese medicinal herb honeysuckle, is a natural antioxidant and serves as anti-inflammatory, anti-tumor, anti-mutagenic and anti-carcinogenic agent. An efficient preparative separation process of chlorogenic acid from honeysuckle crude extracts has been developed in the present study. HPD-850 resin offers the best adsorption capacity, and adsorption and desorption ratios for chlorogenic acid among the nine macroporous resins tested, and its adsorption rate at 25 degrees C fit best to the Langmuir isotherm. The adsorption capacity of HPD-850 resin was found to depend strongly on the pH value of the initial adsorption solution. The dynamic adsorption and desorption experiments have been carried out on a HPD-850 resin packed column to optimize the separation process of chlorogenic acid from honeysuckle crude extracts. After one run treatment with HPD-850 resin, the chlorogenic acid content in the final product was increased 4.46-fold from 11.2% to 50.0%, with a recovery yield of 87.9%. The preparative separation of chlorogenic acid can be easily and efficiently achieved via adsorption and desorption on HPD-850 resin, and the method developed will provide a potential approach for large-scale separation and purification of chlorogenic acid for its wide pharmaceutical use.

Melatonin improves the survival of cryopreserved callus of Rhodiola crenulata

Abstract:  An important aspect of the function of melatonin seems to be the mediation of stress caused by environmental and chemical factors. In the cryopreservation process, environmental changes including osmotic injury, desiccation, and low temperature can impose a series of stresses on plants. In this study, we evaluated the role of melatonin in stress protection during the process of cryopreservation using callus of an endangered plant species Rhodiola crenulata. The survival rate of the cryopreserved callus significantly increased when the callus was pretreated for 5 days with 0.1 μm melatonin prior to freezing in liquid nitrogen. Analysis of antioxidative activity following the pretreatment of callus with 0.1 μm melatonin showed a significant reduction in malondialdehyde production during various steps of cryopreservation. Enhanced peroxidase and catalase activity was observed in the callus after pretreatment with 0.1 μm melatonin compared to the control. These observations provide new evidence of the antioxidant/anti‐stress function of melatonin, and it is the first report of its potential application in the preservation of elite endangered germplasm through the process of cryopreservation.

Efficient harvesting of marine microalgae Nannochloropsis maritima using magnetic nanoparticles

An efficient magnetic separation technology using Fe(3)O(4) nanoparticles was developed for harvesting marine microalgae Nannochloropsis maritima from culture broth. Recovery capacity of these nanoparticles was affected by microalgal growth phase and reached the peak value when the microalgal growth reached its maximal biomass after 18 days. The recovery efficiency of microalgal cells from the culture medium reached more than 95% at the particle dosage of 120 mg/L within 4 min. Electrostatic attraction at acidic pH and cell aggregation under neutral and alkaline conditions was beneficial for harvesting the algal cells. Higher operation temperature resulted in higher adsorption capacity of these nanoparticles for microalgawl cells. Reuse of the culture medium obtained from magnetic separation gave similar biomass production in comparison with that from centrifugation separation after 5 recycles. Together with these results provide a great potential in high-efficient and economical harvesting of tiny marine microalgae using magnetic separation technology in practice.

Enhanced phenol degradation in coking wastewater by immobilized laccase on magnetic mesoporous silica nanoparticles in a magnetically stabilized fluidized bed

The immobilized laccase on magnetic mesoporous silica nanoparticles has been developed for efficient phenol degradation. The degradation rate of phenol by the immobilized laccase was 2-fold higher than that of the free laccase, and the immobilized laccase retained 71.3% of its initial degradation ability after 10 successive batch treatments of coking wastewater. The phenol degradation in the coking wastewater was enhanced in a continuous treatment process by the immobilized laccase in a magnetically stabilized fluidized bed (MSFB) because of good mixing and mass transfer. The degradation rate of phenol maintained more than 99% at a flow rate of less than 450mLh(-1) and decreased slowly to 91.5% after 40h of the continuous operation in the MSFB. The present work indicated that the immobilized laccase on magnetic mesoporous supports together with the MSFB provided a promising avenue for the continuous enzymatic degradation of phenolic compounds in industrial wastewater.

Production of Mouse Interleukin-12 Is Greater in Tobacco Hairy Roots Grown in a Mist Reactor Than in an Airlift Reactor

We compared the growth and productivity of a tobacco line of hairy roots that produces murine interleukin 12 (mIL‐12) grown in three different culture systems: shake flasks, an airlift reactor, and a scalable mist reactor. Of the total mIL‐12 produced by cultures grown in shake flasks (∼434.8 µg L−1), almost 21% was recovered from the medium. In contrast to roots harvested from shake flasks and the mist reactor, roots were not uniformly distributed in the airlift reactor. Roots formed a dense ring around the wall of the reactor and surrounding the central rising column of fine aeration bubbles. Root quality was also better in both the shake flasks and mist reactor than in the airlift reactor. There were more pockets of dark roots in the airlift reactor suggesting some of the roots were nutrient starved. Although the best root growth (7 g DW L−1) was in the shake flasks, both reactors produced about the same, but less dry mass, nearly 5 g DW L−1. Total mIL‐12 concentration was highest in the mist reactor at 5.3 µg g−1 FW, but productivity, 31 µg g−1 FW day−1 was highest in shake flasks. Roots grown in the mist reactor produced about 49.5% more mIL‐12 than roots grown in the airlift reactor. Protease activity in the media increased steadily during culture of the roots in all three systems. The comparisons of protease activity, protein and mIL‐12 levels done in the shake flask system suggest that the increase in proteases associated with progression into stationary phase is most detrimental to mIL‐12 concentration. This is the first description of the design and operation of a scalable version of a mist bioreactor that uses a plastic bag. This also the first report of reasonable production levels of functional mIL‐12, or any protein, produced by hairy roots grown in a mist reactor. Results will prove useful for further optimization and scale‐up studies of plant‐produced therapeutic proteins. Biotechnol. Bioeng. 2009;102: 1074–1086.

Light-enhanced caffeic acid derivatives biosynthesis in hairy root cultures of Echinacea purpurea

Light plays an important role in almost all plant developmental processes and provides the fundamental building blocks for growth, development, primary and secondary metabolism. The effects of light on growth rate and caffeic acid derivative (CADs) biosynthesis in hairy root cultures of Echinacea purpurea (Moench) were assessed. Light-grown hairy roots accumulated increased levels of anthocyanins, which became visible in outer cell layer of the cortex as a ring of purple color. The light-grown root cultures also had radially thickened morphology compared with the dark-grown controls. The growth rate and cell viability of the hairy root cultures in light did not show obvious difference in comparison with those in dark. However, biosynthesis of CADs including cichoric acid, caftaric acid, chlorogenic acid and caffeic acid was significantly increased in hairy root cultures grown in the light. The enhanced accumulation of CADs and anthocyanins in E. purpurea hairy root cultures was correlated to an observed light-stimulated activity of phenylalanine ammonium lyase (PAL).

Magnetic Flocculant for High Efficiency Harvesting of Microalgal Cells

Magnetic flocculant was synthesized for the highly efficient recovery of microalgal cells. The highest flocculation was achieved using the magnetic flocculant synthesized with iron oxide and 0.1 mg/mL cationic polyacrylamide (CPAM). This resulted in a recovery efficiency of more than 95% within 10 min using a dosage of 25 mg/L for Botryococcus braunii and 120 mg/L for Chlorella ellipsoidea. For both species, the adsorption isotherm data fit the Freundlich model better than the Langmuir model, indicating that the adsorption process was a heterogeneous multilayer. The maximum adsorption capacity was 114.8 and 21.4 mg dry cells/mg-particles at pH 7 for B. braunii and C. ellipsoidea, respectively. The primary flocculation mechanism was bridging, which was assisted by the electrostatic interactions between the microalgal cells and the magnetic flocculant under acidic conditions. These results provide new opportunities and challenges for understanding and improving the harvesting of microalgae using magnetic separation.