碩士 / 國立臺灣海洋大學 / 食品科學系 / 107 / This study used enzyme-assisted extraction on phycoerythrin (PE) and phycocyanin (PC) from Porphyra dentata, and extracted Porphyra residues by lactic acid fermentation to produce biopreservative phenyllactic acid (Ph-LA). Using 0.5% (w/v) Porphyra powder to induce marine bacteria Pseudomonas vesicularis MA103 and Aeromonas salmonicida MAEF108 to produce curd enzyme, results showed two marine strains could produce better agarose 3.73 U/mL and 1.17 U/mL on 1 day and 2 days, respectively. Used different solvents (dH2O and Na-P buffer) and assisted extraction methods (enzyme, homogenization, and ultrasound) to extract 2% (w/v) dry Porphyra dentata’s or 2% (w/v) Porphyra powder’s phycoerythrin (PE) and phycocyanin (PC) on 0–72 hr. 2% (w/v) Porphyra powder added 15% (v/v) MA103 and 15% (v/v) MAEF108 induced crude enzyme solution in buffer after reaction 24 hr could gained 0.23 mg/mL of PE and PC (11.5 mg/g PE and PC yield), the PE and PC yields increased by 77% and 109% compared to the dry Porphyra hydrolyzed under dH2O for 24 hr. Selected the best process for extracting PE and PC from Porphyra dentata by 100 kDa Ultrafiltration (UF) distinguish, preliminary evaluation of this process will produce 12.9% UF eluate (< 100 kDa) and 35.1% Porphyra residues, so used that residues to produce Ph-LA by biorefinery concept. Further, 1.56 L Porphyra enzymatic hydrolysates differentiated by 100 kDa UF to 156 mL (> 100 kDa solution), 1.70 mg/mL PE and 1.90 mg/mL PC were available, their purity is 1.04 and 0.74 by A560/A280 and A615/A280, respectively. Then, PE and PC were distinguished by Fast protein liquid chromatography (FPLC). FPLC chromatogram detected 3 major peaks on 560 nm, collected and analyzed that purity index are 4.25 and 0.25, 1.96 and 3.28, 2.26 and 0.35, respectively. Laboratory separated four producing Ph-LA lactic acid bacteria (LAB) strains Lb. plantarum KP3, Lb. plantarum KP4, Leuconostoc mesenteroides K8, and Lb. paracasei subsp. paracasei DP2 cultivate in MRS broth at 37oC by 0–72 hr, strains KP3 and KP4 could produced higher Ph-LA concentrations about 0.09 mg/mL at 24 hr and 36 hr, respectively. Therefore, strains KP3 and KP4 were used in Porphyra residues to produce biopreservative Ph-LA at 37oC. 2% (w/v) Porphyra residues (0.4 g) + 8.21% (w/v) UF eluate + 1.36% (w/v) phenylalanine (Phe) and 0.5% (w/v) yeast extract in 20 broth inoculated 1% lactic acid bacteria strain KP3 fermented at 37oC for 120 hr. The Ph-LA content of fermented broth was 1.86 mg, and the conversion rate from Phe to Ph-LA is 40.46%. Lactic acid bacteria strain KP3 incubated in 8% Porphyra residues + 1.36% Phe and 0.5% yeast extract after 2,000 U commercial cellulase hydrolyzed in 12 hr at 20 mL, and fermented 120 hr could produced 4.58 mg Ph-LA, was 2.5 times concentration of Ph-LA in the MRS group, and the fermentation broth inhibitory activity of angiotensin-converting enzyme (ACE) was 93.8% at a concentration of 3.13%. The ACE inhibitory activities of R-PE and R-PC prepared by the purification of this experiment were 54.8% and 50.8%, respectively. Preliminary evaluation the benefits of this biorefinery concept, enzymatic hydrolysate volume scale up to 1.56 L (24 g Porphyra powder) can produce 32.87 mg R-PE, 49.03 mg R-PC and 37.56 mg Ph-LA, and that in 100 g Porphyra powder (120 NTD) could output products worth 2,455,000 NTD.
| Identifer | oai:union.ndltd.org:TW/107NTOU5253067 |
| Date | January 2019 |
| Creators | Chen, Wei-Chen, 陳韋辰 |
| Contributors | Pan, Chorng-Liang, 潘崇良 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 95 |
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