scholarly journals Protoplast isolation from Dictyopteris pacifica and Scytosiphon lomentaria, using a simple commercial enzyme preparation

Author(s):  
Jose Avila-Peltroche ◽  
Boo Yeon Won ◽  
Tae Oh Cho

Abstract Background Protoplasts (i.e., naked plant cells) can be used for in vitro manipulations and genetic improvement in cultivars with economic value. During the last decade, protoplast research in economic brown algae has been scarce, and it is usually hampered by the use of non-commercial enzymes or crude extracts for isolating protoplasts. Dictyopteris pacifica is part of a brown algal genus well known by its wide chemical diversity and biological properties. Scytosiphon lomentaria is an edible brown seaweed with antioxidant, antitumor, and antiviral properties. So far, there are no protoplast isolation protocols using commercial enzymes for these two economic brown algae. In this study, we obtained protoplasts from cultured samples of D. pacifica and S. lomentaria using commercially available enzymes. Additionally, we investigated the effects of Driselase inclusion and Ca-chelation pre-treatment on protoplast yields in order to optimize the conditions for protoplast preparations. Results Protoplasts were isolated from Dictyopteris pacifica and Scytosiphon lomentaria using the commercially available Cellulase Onozuka RS (1%) and Alginate lyase (4 U mL−1), and short incubation time (4 h). Driselase did not show significant effects on protoplast production in both species. Ca-chelation pre-treatment only increased the number of protoplasts in D. pacifica. Under optimal conditions, the protoplast yields from D. pacifica and S. lomentaria were 4.83 ± 2.08 and 74.64 ± 32.49 × 106 protoplasts g−1 fresh weight, respectively. The values obtained for S. lomentaria were 2–3 orders of magnitude higher than previously reported. Conclusions Our results show that high protoplast yields can be obtained from D. pacifica and S. lomentaria using a simple mixture of commercial enzymes (Cellulase RS and Alginate lyase) and short incubation time (4 h). This work also represents the first report of protoplast isolation in D. pacifica. The method proposed here can help to expand protoplast technology in more brown algal species.

Marine Drugs ◽  
2021 ◽  
Vol 19 (11) ◽  
pp. 628
Author(s):  
Shu-Kun Gao ◽  
Rui Yin ◽  
Xiao-Chen Wang ◽  
Hui-Ning Jiang ◽  
Xiao-Xiao Liu ◽  
...  

Alginate, the most abundant polysaccharides of brown algae, consists of various proportions of uronic acid epimers α-L-guluronic acid (G) and β-D-mannuronic acid (M). Alginate oligosaccharides (AOs), the degradation products of alginates, exhibit excellent bioactivities and a great potential for broad applications in pharmaceutical fields. Alginate lyases can degrade alginate to functional AOs with unsaturated bonds or monosaccharides, which can facilitate the biorefinery of brown algae. On account of the increasing applications of AOs and biorefinery of brown algae, there is a scientific need to explore the important aspects of alginate lyase, such as catalytic mechanism, structure, and property. This review covers fundamental aspects and recent developments in basic information, structural characteristics, the structure–substrate specificity or catalytic efficiency relationship, property, molecular modification, and applications. To meet the needs of biorefinery systems of a broad array of biochemical products, alginate lyases with special properties, such as salt-activated, wide pH adaptation range, and cold adaptation are outlined. Withal, various challenges in alginate lyase research are traced out, and future directions, specifically on the molecular biology part of alginate lyases, are delineated to further widen the horizon of these exceptional alginate lyases.


1996 ◽  
Vol 39 (1-6) ◽  
Author(s):  
K. Stefanov ◽  
V. Bankova ◽  
St. Dimitrova-Konaklieva ◽  
R. Aldinova ◽  
K. Dimitrov ◽  
...  

1986 ◽  
Vol 32 (10) ◽  
pp. 1966-1968 ◽  
Author(s):  
K V Waite ◽  
G F Maberly ◽  
G Ma ◽  
C J Eastman

Abstract We adapted a commercial immunoradiometric assay (IRMA) to measure thyrotropin in filter-paper blood spots. Two 3-mm blood spots are used for each standard and sample. These are incubated for 2 h with radiolabeled antibody and for 30 min with magnetic antibody, followed by a 10-min separation procedure. Assay sensitivity is 6 milli-int. units/L. Coefficients of variation (precision profile of the standard curve) ranged from 4.3 to 9.6%. The coefficient of correlation (r) between thyrotropin concentrations in the blood spots and in serum was 0.93. Pre-elution of the blood spots is necessary for short incubation time. Short incubation time, little need for specialized equipment, the high precision and sensitivity characteristic of IRMA, and ease of collection, transport, and storage of the blood-spot samples make this assay suitable for neonatal hypothyroid screening.


1982 ◽  
Vol 65 (5) ◽  
pp. 1129-1133 ◽  
Author(s):  
William D St John ◽  
Jack R Matches ◽  
Marleen M Wekell

Abstract A simple iron milk medium was used for isolation and enumeration of Clostridium perfringens from soil, sludge, and water samples. The whole milk contained only iron powder as a reducing agent; no other inhibitors were added. The iron milk most probable number (MPN) procedure was compared with 4 plating media: sulfite-polymyxin-sulfadiazine, Shahidi-Ferguson perfringens, tryptose-sulfite- cycloserine (both with and without egg yolk), and tryptone-sulfite-neomycin. The selectivity of the iron milk relies solely on the rapid growth of C. perfringens at 45°C and the stormy fermentation reaction within 18 h. Isolates were confirmed as C. perfringens by standard biochemical tests. The iron milk MPN procedure compared very well with the 4 plating media tested. Selectivity of incubation temperature, short incubation time, and ease of identification by the characteristic stormy fermentation make this method ideal for enumerating C. perfringens from large numbers of samples.


CATENA ◽  
2018 ◽  
Vol 163 ◽  
pp. 354-360 ◽  
Author(s):  
Adriano dos Santos Moura ◽  
Erika Valente de Medeiros ◽  
Jéssica Emanuella da Silva Oliveira ◽  
Rafaela Felix da Franca ◽  
Anderson Dantas Lira ◽  
...  

1992 ◽  
Vol 282 ◽  
Author(s):  
D. Srinivas ◽  
R. Foster ◽  
S. Marcus ◽  
R. Arora ◽  
H. Rebenne

ABSTRACTIn this work, a hydrogen (H2) reduction process has been developed which gives tungsten (W) nucleation on titanium nitride (TiN) adhesion layers with a very short incubation time, eliminating the need for a silane (SiH4reduced seed layer. The nucleation was found to be strongly dependent on the following factors: temperature of the substrate, total pressure in chamber, and gas introduction sequence into the reactor. Theenhanced nucleation rate has been explained based on two competing reactions: dissociation of H2, and formation of titanium subfluorides on the TiN surface.


Foods ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3050
Author(s):  
Mika Immonen ◽  
Julia Myllyviita ◽  
Tuula Sontag-Strohm ◽  
Päivi Myllärinen

The aim of this study was to develop an extraction method to produce highly functional oat protein concentrates. We investigated the possibility of combining enzyme-aided slightly alkaline (pH 8.0) extraction with ultrafiltration and subsequent diafiltration for concentration of the extracted oat proteins. A further aim was to study how the deamidation of oat proteins with protein-glutaminase (PG) improves the solubility of proteins as a function of the following parameters: pH (6.0–9.0), enzyme dosage (4–20 U/g protein), and incubation time (1–4 h) with response surface methodology (RSM). Furthermore, we investigated selected functional properties, such as heat-induced gelation and solubility, of the oat protein concentrates. The chosen parameters for the enzymatic deamidation pre-treatment process by PG were as follows: pH 8.0, dosage 11.0 U/g protein, and an incubation time of 4 h (1 h at native pH and 3 h at pH 8.0). Two oat protein concentrates were produced, non-deamidated and ultrafiltered, and deamidated and ultrafiltered, with protein concentrations of 45.0 and 52.4%, respectively. The solubility of both oat protein concentrates was significantly improved at neutral and slightly alkaline pH compared to the solubility of proteins extracted from the starting material. Additionally, both oat protein concentrates produced equally strong heat-induced gel-like structures at a protein concentration of 10%.


2020 ◽  
Vol 3 (2) ◽  
pp. 98-107
Author(s):  
Galileo E. Araguirang ◽  
Arianne Joyce R. Arizala ◽  
Eden Beth B. Asilo ◽  
Jamie Louise S. Batalon ◽  
Erin B. Bello ◽  
...  

Banana (M. acuminata x balbisiana) is an abundant lignocellulosic waste material in large plantations all over the Philippines, especially in Mindanao, which can be utilized as substrate in producing high-value products like ethanol. To compensate for the low yield based on total weight of substrate due to the high moisture content of banana pseudostem, there is the primary challenge to make the conversion of this lignocellulosic biomass into monomeric sugar and then into ethanol more efficiently in order to achieve yields that would make it cost-competitive. Hence, this study evaluated the effects of solid loading, incubation time and amount of enzyme on yield of reducing sugars in the enzymatic hydrolysis process and attempted to optimize the significant factors by Response Surface Methodology (RSM), specifically using Box-Behnken design. There was significant improvement on the reducing sugar yield of the pretreated banana pseudostem at 20 h incubation time, 15 g solid loading and 0.55 % enzyme concentration. Ethanol production was observed to be higher in the detoxified substrate although biomass was higher for the non-detoxified substrate. As to our knowledge, the present study is the first attempt to produce second generation ethanol using banana pseudostem waste as feedstock in the Philippines.


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