An integrated process to recover a spectrum of bioproducts from fresh seaweeds

Patent metadata

Field	Value
Publication number	US-2016355610-A1
Application number	US-201415109232-A
Country	US
Kind code	A1
Filing date	Dec 30, 2014
Priority date	Dec 30, 2013
Publication date	Dec 8, 2016
Grant date	—

Abstract

An integrated process for the recovery of a spectrum of commercially valuable products such as agar, cellulose, lipids, pigments and a liquid rich in minerals of agricultural importance directly from fresh seaweed without employing any catalyst driven in situ chemical conversions. Also solvents used during lipid extraction were shown to be used for three cycles without affecting the yield and quality of successive products. Furthermore, this new process is highly efficient and utilizes total seaweed raw material without any leftover biomass as solid waste.

First claim

1 . An integrated process to recover a spectrum of bioproducts from fresh seaweeds, wherein the steps comprising:
[a] homogenizing the fresh seaweeds using mixer grinder in phosphate buffer having pH in the range of 6.8 to 7.0 and incubating at a temperature in the range of 4 to 6 degree C. for a time period of 10 to 12 hours followed by centrifugation at 5000 to 7000 rpm at 4 to 10 degree C. for 12 to 15 minutes and separating the supernatant containing pigments and protein from residual mass; [b] precipitating the pigments from supernatant as obtained in step [a] with 30% agarophytes and 40% carrageenophytes of ammonium sulphate followed by centrifugation at 6500 to 8000 rpm at 4 to 10 degree C. for 12 to 15 minutes and dissolving the pelleted pigments in phosphate buffer having pH in the range of 6.8 to 7.0 followed by quantification of the pigments using spectrophotometric method, recording the absorbance using double beam UV-Vis spectrophotometer at 280, 564, 618 and 730 nm and calculating the concentration of pigments R-PE and R-PC according to the equation:
R-PC=0.154(A 618 -A 730 )
R-PE=0.1247((A 564 -A 730 )−0.4583(A 618 -A 730 ));

[c] analyzing the composition of supernatant obtained from step [b] for the presence of various plant nutrient including macro and micro minerals using Inductive Coupled Plasma (ICP) spectroscopy; [d] extracting the total lipids from the residual mass as obtained in step [a] using the solvents chloroform and methanol mixed in a ratio of 1:2 v/v followed by centrifugation at 4000 to 5000 rpm at 4 to 10 degree C. for 10 to 15 minutes and separating the aqueous and greenish organic extract layer from the residual mass; [e] filtering the organic extract as obtained in step [d] and washing with equal volume of milli Q water followed by drying using rotary evaporator and gravimetric quantification of lipids; [f] subjecting the residual mass as obtained in step [d] for agar extraction, if the seaweed is an agarophyte by drying it at a temperature in the range of 60 to 65 degree C. for 50 minute to 1 hr followed by cooking with 1:5 w/v (of initial fresh algal biomass) of distilled water at 115 to 120 degree C. for 1 hr 20 minutes to 1.5 hours in an autoclave and homogenizing the cooked material immediately using mixer grinder followed by centrifugation at 6000 to 7000 rpm at 40 to 60 degree C. for 4 to 6 minutes to obtain the supernatant and residual mass; [g] allowing the supernatant as obtained in step [f] to form a gel at temperature in the range of 25 to 30 degree C. followed by freezing at minus 15 to minus 20 degree C. for 12 to 15 hours and thawing to obtain the native agar which is dried at 60 to 65 degree C. for 10 to 12 hours to obtain the dry agar; [h] subjecting the residual mass as obtained in step [d] for carrageenan extraction, if the seaweed is a carrageenophyte by drying it at a temperature in the range of 60 to 65 degree C. for 50 minutes to 1 hr followed by cooking with 8% KOH at 70 to 72 degree C. for 1 hr 45 minutes to 2 hours, filtering the contents through muslin cloth and washing the filtrate with tap water till neutrality followed by cooking it with 100 ml water at 75 to 78 degree C. for 40 to 45 minutes; [i] homogenizing the cooked material as obtained in step [h] and centrifuging at 6000 to 7000 rpm for 3 to 5 minutes to obtain the supernatant and residual mass, precipitating the supernatant in chilled iso-propanol (1:2 v/v) and drying the precipitate at 60 to 65 degree C. for 4 to 6 hours to obtain carrageenan; [j] characterizing the agar and carrageenan obtained in steps [g] and [i] respectively using Fourier transformation infrared (FT-IR) spectroscopy and measuring the gel strength of agar and carrageenan following standard methods; [k] drying the residual mass as obtained in steps [f] and [i] and soaking in acetate buffer of pH 4.5 to 5 containing 36% NaClO 2 (w/w) for bleaching at 60 to 65 degree C. for 7 to 8 hours, washing of bleached material with water to neutralize it and treating the washed material with 0.5 M NaOH solution at 60 to 65 degree C. for 10 to 12 hours followed by washing with water till neutrality; [l] drying the neutralized material as obtained in step [k] and treating with 5% v/v of hydrochloric acid followed by heating till boiling, incubating at temperature of 25 to 30 degree C. for 10 to 14 hours and washing the slurry with water till neutrality followed by drying to get cellulose; [m] hydrolyzing the cellulose as obtained in step [l] with commercial cellulase in sodium acetate buffer of pH 4.8 and incubating for 36 hrs at 45 degree C. on an orbital shaker, measuring the reducing sugar spectrophotometrically using 3, 5-dinitrosalisylic acid method; [n] enriching the hydrolysate as obtained in step [m] with 5 g/L peptone and 3 g/L yeast extract followed by sterilization in an autoclave, inoculating the culture of Saccharomyces cerevisiae , and incubating it for 12 hrs at 28±2 degree C. on an orbital shaker to obtain ethanol; analyzing the ethanol yield and residual reducing sugars by GC-MS and DNS method, respectively.

2 . A process as claimed in claim 1 , wherein, the yields of the products are comparable with those of obtained individually following conventional method.

3 . The process as claimed in claim 1 wherein, pretreatment is not required for agar extraction oppose to the conventional extraction methods.

4 . The process as claimed in claim 1 wherein, it enables to produce agar with high gel strength without alkali treatment as conventionally done.

5 . The process as claimed in claim 1 wherein, the gel strength of phycocolloid (agar) obtained is superior to that of the same obtained from conventional agar extraction and shows 1.5 to 2.9 fold increase in gel strength.

6 . The process as claimed in claim 1 wherein, the produced agar for different species ( Gelidiella acerosa, Gelidium pusillum and Gracilaria dura ) are suitable for microbiological application with equal or lower concentrations.

7 . The process as claimed in claim 1 wherein, the produced agar shows gel strength in the range of 500-1200 g/cm 2 .

8 . The process as claimed in claim 1 wherein, after recovery of multiple products sequentially yielded 10-25% left over residue which is then used as a feedstock for cellulose extraction.

9 . The process as claimed in claim 1 wherein, the residual mass minimizes the use of chemicals up to 75 to 90% for cellulose extraction.

10 . The process as claimed in claim 1 wherein, cellulose produced without any additional decoloration and defatting oppose to conventional methods.

11 . The process as claimed in claim 1 wherein, the liquid obtained after salting out of pigment contained high amount ammonium sulphate in addition to good quantity of seaweeds based macro- and micro-minerals.

12 . The process as claimed in claim 1 wherein, the solvents used during lipid extraction were shown to be reusable for three cycles without compromising on the yield and quality of successive products.

Assignees

• Council Scient Ind Res

Inventors

• Reddy Chennur Radhakrishna
• Baghel Ravi Singh
• KUMARI Nitin
• Kumari Puja
• Gupta Vishal
• Prasad Kamlesh
• Meena Ramavatar

Classifications

• C08B37/0039

  Agar; Agarose, i.e. D-galactose, 3,6-anhydro-D-galactose, methylated, sulfated, e.g. from the red algae Gelidium and Gracilaria; Agaropectin; Derivatives thereof, e.g. Sepharose, i.e. crosslinked agarose · CPC title

• C12P7/065

  with microorganisms other than yeasts · CPC title

• C11B1/10

  by extracting · CPC title

• C12P19/14

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C08B37/0042

  Carragenan or carragen, i.e. D-galactose and 3,6-anhydro-D-galactose, both partially sulfated, e.g. from red algae Chondrus crispus or Gigantia stellata; kappa-Carragenan; iota-Carragenan; lambda-Carragenan; Derivatives thereof · CPC title