• Journal of Microbiology and Biotechnology
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• 제32권12호
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• pp.1599-1604
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• 2022

Storage stability of freeze-dried lactic acid bacteria is a critical factor for their cost-effectiveness. Long-term storage of lactic acid bacteria enables microbial industry to reduce distribution costs. Herein, we investigated the effect of cold adaptation under supercooling conditions at -5℃ on the viability of Leuconostoc mesenteroides WiKim32 during the freeze-drying process and subsequent storage. Cold adaptation increased the thickness of exopolysaccharides (EPS) and improved the viability of freeze-dried Leu. mesenteroides WiKim32. Compared to non-adapted cells, cold-adapted cells showed a 35.4% increase in EPS thickness under supercooling conditions. The viability of EPS-hydrolyzed cells was lower than that of untreated cells, implying that EPS plays a role in protection during the freeze-drying process. Cold adaptation increased the storage stability of freeze-dried Leu. mesenteroides WiKim32. Fifty-six days after storage, the highest viability (71.3%) was achieved with cold adaptation at -5℃. When EPS-containing broth was added prior to the freeze-drying process, the viability further increased to 82.7%. These results imply that cold adaptation by supercooling pretreatment would be a good strategy for the long-term storage of Leu. mesenteroides WiKim32.

• Journal of Microbiology and Biotechnology
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• 제12권2호
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• pp.349-353
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• 2002

Two recombinant bacteria containing luxAB showed an increased tolerance to stresses associated with lyophilization, when the cells were freeze-dried in the presence of trehalose. In the case of a recombinant, UV2, only $2.5\%$ of the original bioluminescence and $2.7\%$ of the cell viability were restored after 4 h of freeze-drying without trehalose, which implies that the cells were heavily damaged during the dehydration. To improve these losses, trehalose was added before freeze-drying using different modes. Trehalose increased the bioluminescence and the viability of freeze-dried UV2 under all conditions tested, and it was also observed that the addition of trehalose to the cultures (final concentration of 0.08 M) for 15 min before the freeze-drying resulted in the restoration of $45\%$ of the original bioluminescence and $50\%$ of the cell viability. Trehalose also showed a similar efficacy with the other luminescent recombinant, YH9. Therefore, it was tentatively concluded that trehalose played a role as a protective agent in the freeze-drying of bacterial cells.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권3호
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• pp.202-206
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• 2000

The genetically-engineered Escherichia coli strain, DPD2540, which contains a fabA:::luxCDAbefusion gene, gives a bioluminescent output when membrane fatty acid synthesis is needed. For more pactical application of this strain in the filed as biosensor, freezedrying was adopted. A 12% surcrose solution with Luria-Bertani (LB) broth, as determined by the viability after freeze-drying, was found to be most most effective composition for lyophilization solution among various compositions testitons tested. Rapid freezing with liquid nitrogen also gave the best viability after freeze-drying as compared to samples frozen at-7$0^{\circ}C$ and -2$0^{\circ}C$. The biosensing activities of the cells showed a greater sensitivity when the cells from the expontial phase were freeze-dried. Finally, the optimum temperature for use of the freeze-dried cells in the biodencor field was determined.

• 대한의생명과학회지
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• 제9권4호
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• pp.195-201
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• 2003

Aromatic hydrocarbons are of major concern among genotoxic chemicals due to their toxicity and persistence. Some microorganisms can utilize aromatic hydrocarbons as carbon and energy sources by inducing expression of catabolic operon(s). The XylR regulatory protein activates transcription of the catabolic enzymes to degrade BTEX (benzene, toluene, ethylbenzene, and xylene) from its cognate promoters, Pu and Ps upon exposure of the cells to the aromatic hydrocarbons. The activity of XylR on the promoters was previously monitored using luciferase luc reporter system. The xylR, its promoter Pr and the promoter Po for the phenolic compound catabolic operon were introduced upstream of firefly luciferase luc in the pGL3b vector to generate about 7.1 kb of pXRBTEX. Here E. coli harboring the plasmid was freeze-dried under various conditions to fin,d optimal conditions for storage and transport. The cell viability and luciferase activity were maintained better, when the cells were freeze-dried at -7$0^{\circ}C$ in the addition of the 10% skim milk or 12% sucrose. However, coaddition of protectants such as 10% skim milk plus 10% glucose or 12% sucrose plus 10% glucose, resulted in much better viability and bioluminescence activity compared with the effect of single addition of each protectant. In addition, it was shown that the freeze-dried cells maintained almost intact bioluminescent activities and cell viability for at least 1 week after freeze-drying. This work demonstrated that the properly freeze-dried recombinant bacterial cells could be utilized as a whole-cell biosensor for simple and rapid monitoring of BTEX in the environment.

• Clinical and Experimental Reproductive Medicine
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• 제51권1호
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• pp.42-47
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• 2024

Objective: This study evaluated the effects of temperature and storage time on the quality and DNA integrity of freeze-dried sperm from individuals with normozoospermia. Methods: Normal sperm samples from 15 men aged 24 to 40 years were studied. Each sample was divided into six groups: fresh, freezing (frozen in liquid nitrogen), freeze-dried then preserved at room temperature for 1 month (FD-1m-RT), freeze-dried then preserved at room temperature for 2 months (FD-2m-RT), freeze-dried then preserved at 4 ℃ for 1 month (FD-1m-4 ℃), and freeze-dried then preserved at 4 ℃ for 2 months (FD-2m-4 ℃). The morphology, progressive motility, vitality, and DNA integrity of the sperm were evaluated in all groups. Results: In all freeze-dried groups, sperm cells were immotile after rehydration. The freeze-dried groups also showed significantly less sperm vitality than the fresh and frozen groups. Significantly more morphological sperm abnormalities were found in the freeze-dried groups, but freeze-drying did not lead to a significantly higher DNA fragmentation index (DFI). The DFI was significantly higher in the FD-2m-RT group than in the other freeze-dried groups. Conclusion: The freeze-drying method preserved the integrity of sperm DNA. The temperature and duration of storage were also identified as factors that influenced the DFI. Accordingly, more research is needed on ways to improve sperm quality in the freeze-drying process.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1428-1440
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• 2017

Phomopsis sp. XP-8 (an endophytic fungus) was previously found to produce pinoresinol diglucoside (PDG), a major antihypertensive compound of Tu-Chung (the bark of Eucommia ulmoides Oliv.), which is widely used in Chinese traditional medicines. In the present study, two bioconversion systems were developed for the production of PDG in Tris-HCl buffer containing glucose and Phomopsis sp. XP-8 cells (both resting and freeze-dried). When other factors remained unchanged, the bioconversion time, glucose concentration, cell ages, cell dosage, pH, temperature, and stirring speed influenced PDG production in a similar and decreasing manner after an initial increase with increasing levels for each factor. Considering the simultaneous change of various factors, the optimal conditions for PDG production were established as 70 g/l cells (8-day-old), 14 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing resting cells, and 3.87 g/l cells, 14.67 g/l glucose, $28^{\circ}C$, pH 7.5, and 180 rpm for systems employing freeze-dried cells. The systems employing freeze-dried cells showed lower peak PDG production ($110.28{\mu}g/l$), but at a much shorter time (12.65 h) compared with resting cells (23.62 mg/l, 91.5 h). The specific PDG production levels were 1.92 and $24{\mu}g$ per gram cells per gram glucose for freeze-dried cells and resting cells, respectively. Both systems indicated a new and potentially efficient way to produce PDG independent of microbial cell growth.

• 한국식품과학회지
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• 제30권6호
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• pp.1448-1455
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• 1998

본 연구에서는 과즙-우유 혼합 기질로 만들어진 발효유를 동결건조하고, 동결건조 전과 후의 생균수, pH의 변화, 관능성 및 휘발성 향기 성분의 변화를 조사하였다. 동결 또는 동결건조에 의하여 발효유의 pH는 거의 변화가 없었으나, 생균수는 동결, 특히 동결건조 도중에 급격히 감소하였는데, 동결 전의 균수를 100%로 했을 때, 동결 후의 생존율은 $64.5{\sim}85.2%$, 동결건조 후의 생존율은 $10.0{\sim}21.1%$였다. 사과쥬스-우유의 혼합 비율이 15:35인 기질로 만든 발효유와 동일한 발효유를 동결건조-환원한 시료의 관능성을 비교했을 때, 동결건조 전의 시료가 후의 시료보다 관능성이 우수하며, 이와 같은 경향은 포도쥬스-우유 혼합 시료의 경우에 보다 현저하였다. 동결건조 전과 후의 모든 시료에서 ethanol, diacetyl, butanol, acetoin이 검출되었고, 포도쥬스 함량이 높은 시료에서는 acetone도 검출되었으며, 동결건조에 의하여 모든 시료의 휘발성 향기 성분의 함량이 감소하는 경향을 보였다. 본 실험에서 검출된 향기 성분 가운데 젖산균 발효에 의하여 생성된 향기 성분은 ethanol, diacetyl, acetoin 이었다.

• 한국식품과학회지
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• 제31권5호
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• pp.1337-1344
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• 1999

본 연구에서는 난백분말과 카제인 기질에 생육촉진물질을 첨가하여 만든 젖산균 발효식품의 동결건조에 의한 젖산균 생균수의 변화, 관능성의 변화, 경도, 휘발성 향기성분의 변화를 관찰하였다. 동결 또는 동결건조에 의하여 젖산균 발효식품의 pH 거의 변화가 없으나, 생균수는 동결, 특히 동결건조 도중에 급격히 감소하였고, 동결 전의 균수를 I00%로 했을 때, 동결 후의 생존율을 $72.0{\sim}82.4%$, 동결건조 후의 생존율은 $10.0{\sim}20.4%$였다. 동결건조 전 시료와 동결건조-환원된 시료의 관능성을 비교하여 보면, 대체적으로 동결건조 전의 시료가 동결건조-환원된 시료보다 관능성이 우수하였으나, 동결건조시킨 고체상의 발효유 및 젖산균 발효식품도 환원성이 뛰어나고 기호도가 양호하였다. 동결건조 전의 젖산균 발효식품과 동결건조-환원된 젖산균 발효식품의 휘발성 향기 성분의 변화를 보면, 동결건조에 의하여 모든 시료의 휘발성 향기 성분의 함량이 감소하는 경향을 보였고 그 감소 정도는 시료 별로 차이가 있었다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제13권3호
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• pp.265-277
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• 1991

In this study, the healing changes of the implanted bone and its surrounding tissues were examined on the histopathologic basis following implantation of the freeze - dried and radiation - sterilized allogeneic bone in Rectus abdominicus of the rat. This study was performed to see the tissue recations after implantation of the freeze - dried and radiation - sterilized allogeneic bone and whether osteogenesis or osteo - induction or osteo - conduction is happened. And the results were as follows : 1. The shape of the implanted allogeneic bone of the 1, 2 - week group specimen was similar to that of normal bone in light - microscopic finding and the atrophy of cellular organells was found in trans - mission electron - microscopic finding. 2. The implanted allogeneic bone was surrounded with the dense fibroconnective tissues, and infiltration of the chronic inflammatory cells gradually became increased. 3. Hyaline degeneration was observed in the surrounding tissue at the 3, 4, 6 - week group specimen. 4. Light - microscopically the resorption of implanted bone became prominent after 4 - week group and the necrosis of allogeneic bone implant became severe with loss of cell components in lacuna. 5. Electron - microscopically, the osteoclast - like cells ere fond after, 2 - week group. It is summarized that the osteo - conduction potential of the bone is remained just after implanting the freeze - dried and radiation - sterilized allogeneic bone on Rectus abdominicus of the rat, but gradually it disappeared with the gradual increse of chronic inflammatory reaction and osteoclastic activity. So it is suggested that the antigenicity of the freeze - dried and radiation - sterilized bone is remained and it has little osteo - conductive activity when it is implanted in the muscle.

• 한국식품과학회지
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• 제43권4호
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• pp.509-512
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• 2011

동결건조 쌀죽의 품질특성에 미치는 동결속도의 영향을 조사하기 위하여 쌀죽을 -20, -40, -70$^{\circ}C$의 동결 온도에서 동결시킨 후 동결건조시킨 제품을 제조하여 이들 제품의 미세구조, 물리적 강도, 조직감 및 재수화율을 조사하였다. 전자주사현미경을 통한 이들 제품의 미세구조를 관찰한 결과 완만동결(-20$^{\circ}C$)시킨 것이 급속동결(-70$^{\circ}C$)에 비해 공극의 크기가 크고 그 수가 적었다. 반면에 급속동결시킨 제품은 보다 조직이 치밀하고 물리적인 강도가 유의적(p<0.05)으로 큰 값을 나타냈다. 수화복원율은 -20$^{\circ}C$에서 동결시킨 제품이 -40, -70$^{\circ}C$에서 동결시킨 제품보다 높았다. 따라서 품질이 우수하고 사용이 간편한 동결건조 쌀죽을 제조하기 위해서는 최적의 동결 온도의 선택이 필수적임을 알 수 있었다.