ACHIEVEMENT OF ZERO EMISSIONS BY THE BIOCONVERSION OF FISHERY WASTES INTO FERTILIZER - Fertilizers: Components, Uses in Agriculture and Environmental Impacts - page 72

positive rods, which measured 0.5-1.2 μm in width and 2-4 μm in length, and formed
endospores. The colors of the colonies that formed on Porphyra powder, agar and
carrageenan media were orange, pink and white, respectively. Because this strain
demonstrated distinct degradation abilities for both agar and carrageenan, we have applied for
a Korean patent (No.: 10-2012-0024992) after depositing this strain into KACC as
KACC91705P. The pure culture was maintained on a 1.5% nutrient agar plate at 4 °C until
use and transferred to a fresh agar plate every month. Periodically, the potential degrading
ability of this strain was checked on both agar and carrageenan agar by the plate assay.
Brown Seaweed
The bacterial strain that was used for the biodegradation of brown seaweed
polysaccharides was Microbacterium oxydans (GenBank Accession No.: HQ113206.1),
which were isolated from silt and sandbar locations in a coastal area near Busan (Korea),
where brown seaweed often drifts and accumulates. This strain formed a transparent ring (on
alginate agar) or a yellow-colored ring (on laminarin agar) around each colony according to
the plate assay (Gacesa & Wusteman, 1990; Lee & Chang, 1995), indicating that this strain
possesses high alginate lyase and laminarinase activities. Based on microscopic observations,
M. oxydans was extremely motile in the vegetative state and displayed Gram-positive rods,
which measured 0.4-0.8 µm in width and 1.0-2.0 µm in length. The strain occurred both
singly or in random groups, was catalase-positive and did not form endospores. Because this
novel strain exhibits the distinct ability to degrade both alginate and laminarin, we have
applied for a Korean patent (No.: 10-2011-0097411) after depositing this strain into KACC as
KACC 91657P. The pure culture was maintained on a 1.5% nutrient agar plate at 4 °C until
use and transferred to a fresh agar plate every month. Periodically, the potential degrading
ability of this strain was checked on both alginate and laminarin agar by the plate assay.
Culture medium
Green Seaweed
A fresh colony of B. licheniformis TK3-Y from a nutrient agar plate was transferred with
a platinum loop to a 10-ml tube containing 5 ml Ulva culture medium (10 g l -1 Ulva powder,
5 g l -1 yeast extract, 2.4 g l -1 NH 4 Cl, 2 g l -1 KH 2 PO 4 , and 0.5 g l -1 NaCl). The Ulva powder
(mixture of U. pertusa and U. lactuca ) was purchased from a local market and ground using a
porcelain mortar and pestle. The final samples were sieved to achieve particle size
homogeneity using a 100-mesh sieve. The ground powder was sonicated for 1 h after
treatment with 1% (v v -1 ) H 2 SO 4 at 121 °C for 15 min to increase its solubility. The pH of the
culture medium was adjusted to 6 before autoclaving, and the culture medium was sterilized
at 121°C for 15 min. After inoculation, the tube was incubated in a rotary shaking incubator
at 50 °C and 180 rpm for 3 d until cells reached a late-log phase. A 5% (v v -1 ) inoculum from
this tube was then used to inoculate 100 ml of the Ulva culture medium in a 250 ml
Erlenmeyer flask, and the flask was incubated for 2 d. Then, wet cells from the flask were
collected by centrifugation at 8000 rpm, and the harvested cells were used as an inoculum for
the biodegradation experiment.