Trehalose와 당류가 냉동요구르트의 저장 중 유산균 생존율과 품질특성에 미치는 영향

우성호,주진우,윤원병,김거유 한국산업식품공학회 2010 산업 식품공학 Vol.14 No.1

본 연구에서는 냉동요구르트 제조 시 trehalose와 sorbitol 첨가가 유산균의 동결변성방지 및 향기성분의 변화에 미치는 영향을 조사하고자 실시하였다. 요구르트를 L. bulgaricus와 S. thermophilus 단독균주 및 혼합균주로 발효시켰을 때 처리구 모두 trehalose와 sorbitol 첨가에 의한 발효과정 중 유산균의 증식효과는 나타나지 않았다. 요구르트를 6주간 냉동저장하는 동안 trehalose와 sorbitol 첨가에 의한 모든 처리구에서 유산균수의 차이가 없었으며, MRS 액상배지에 trehalose와 sorbitol을 각각 2%, 5% 첨가하여 6주간 냉동저장하였을 때는 5% trehalose 처리구의 동결변성방지 효과가 가장 우수한 것으로 나타났다. 냉동저장 기간 동안 고형분 함량이 12%인 처리구와 20%인 처리구의 생균수는 큰 차이가 나타나지 않아 고형분 함량 차이에 의한 trehalose와 sorbitol 효과에는 차이가 없는 결과를 나타내었다. 요구르트의 동결과 해동을 반복하였을 경우 trehalose와 sorbitol 모두 2% 첨가구보다 5% 첨가구가 우수한 동결변성방지 효과를 나타내었다. 전반적으로 Trehalose 첨가구가 sorbitol 첨가구보다 2%, 5% 모두 동결변성방지 효과가 좋은 결과를 나타내었다. Trehalose와 sorbitol을 첨가하여 요구르트를 제조하였을 때 향미성분의 함량은 대조구와 유사하였으며 일반적인 요구르트의 향미를 나타내었다. 냉동저장 중 요구르트 향미성분은 손실되거나, 함량비율의 변화가 발생하지 않았다. 이상의 결과로부터 요구르트의 풍미에는 영향을 미치지 않으면서 냉동저장에 의한 유산균의 동결변성방지제로서의 trehalose첨가가 냉동요구르트의 유산균 안정화에 효과가 있다고 하겠다. The objective of the current study was to determine the cryoprotective effects of trehaolse on lactic acid bacteria in the frozen yoghurt during long-term frozen storage conditions. The frozen yoghurts were prepared using starter culture containing Lactobacillus bulgaricus and Streptococcus thermophilus, as well as trehalose and sorbitol as a cryoprotectant. The viable cell numbers of lactic acid bacteria in frozen yoghurt did not significantly decreased during six weeks frozen storage conditions. The MRS broth, which contains either trehalose or sorbitol, cultured with L. bulgaricus and/or S. thermophilus, and then the cultured medium was kept in the frozen condition for six weeks. The results indicated that lactic acid bacteria viability significantly increased with trehalose addition (2 and 5%) in the media compared to those of control and sorbitol supplement groups. The lactic acid bacteria viability in the yoghurts was examined on the effects of repeated freeze and thaw events. The freeze-thaw resistance of lactic acid bacteria significantly increased with trehalose supplement in the yoghurt. The major volatile aroma compounds (acetaldehyde, acetone, ethanol, diacetyl, and acetoin) in yoghurt were separated and indentified by headspace GC-FID analysis. Distinct flavor components and their ratios are known as important quality factors for yoghurt notes. Trehalose addition to the yoghurt was not influenced these factors during lactic acid fermentation. The results in this study demonstrated that trehalose potentially can be applicable as an effective cryoprotectant for lactic acid bacteria in the frozen yoghurt products.

Trehalose가 빵용 반죽의 Rheology 특성에 미치는 영향

김영자(Young-Ja Kim),이정훈(Jeong-Hoon Lee),정구춘(Koo-Chun Chung),이시경(Si-Kyung Lee) 한국식품과학회 2014 한국식품과학회지 Vol.46 No.3

Trehalose가 밀가루 반죽의 rheology에 미치는 영향을 평가하기 위하여 밀가루 대비 설탕을 6% 첨가한 것을 대조구로 하여 trehalose를 각각 2, 4, 6% 첨가하여 farinograph, viscograph, 반죽의 pH 및 총산도, 반죽의 발효팽창력 등을 분석하였다. Farinograph 분석에서 trehalose 첨가량이 증가함에 따라 반죽의 견도와 반죽형성시간은 증가하였으나, 흡수율, 안정도, 약화도, FQN 등은 감소하였다. Viscograph 분석에서 trehalose 첨가가 호화개시온도, 최고점도온도에는 영향을 주지 못하였으나, 최고점도는 trehalose 6% 첨가구에서 가장 낮았다. Trehalose 첨가량이 증가할수록 최종점도, breakdown, setback 등은 감소하였다. 120분 발효 동안 반죽의 pH 변화는 trehalose 4% 첨가구가 가장 낮았고, 반죽의 총산도는 trehalose 4% 첨가구가 가장 높았다. 반죽의 발효팽창력도 trehalose 4% 첨가구가 다른 시험구에 비해 큰 부피를 나타냈다. 이상과 같이 trehalose 첨가량을 달리한 실험에서 trehalose의 첨가는 빵용 밀가루 반죽의 rheology 특성에 영향을 주어 빵 제조에 첨가 시 빵의 부피가 크고, 부드럽고, 노화가 지연될 것으로 생각된다. This study was conducted to evaluate the effect of trehalose on the rheological properties of bread flour dough. Farinographic and viscographic properties, pH, total titratable acidity (TTA), and fermentation power were analyzed for flour dough rheology. Flour dough containing trehalose showed greater water absorption capacity and longer development time. However the stability, degree of softening, and farinograph quality number (FQN) were lower for the trehalosecontaining flour dough, however, these factors decreased with increasing amounts of trehalose. Trehalose did not affect the beginning of gelatinization temperature and maximum viscosity temperature of flour. The maximum viscosity was the lowest with 6% trehalose, the end of final holding period, breakdown and setback values decreased with increasing amounts of trehalose. Flour dough with 4% trehalose had the lowest pH value for 120 min fermentation at 30℃, and the highest TTA value. Addition of 4% and 6% trehalose showed larger fermentation volume of dough than the control. The results suggested that trehalose positively affected the rheological properties of flour dough such as bread volume, softness, and staling delay.

Trehalose에 의한 Rahnella aquatilis AY2000 균주가 생산하는 항효모성 물질의 활성보호

강민정,이종환,이복규,김광현,Kang, Min-Jung,Lee, Jong-Hwan,Lee, Bok-Kyu,Kim, Kwang-Hyeon 한국미생물·생명공학회 2009 한국미생물·생명공학회지 Vol.25 No.4

AYS(Anti-yeast substance)는 열에 의해 항효모 활성이 쉽게 약화되는 물질로서 trehalose의 사용이 AYS의 열에 대한 불안정성을 어느 정도 극복할 수 있을 것인지를 조사하기 위해 trehalose가 첨가된 AYS와 trehalose가 첨가되지 않은 AYS에 대한 항효모 활성을 비교 측정하였다. 그 결과 trehalose가 첨가된 AYS는 고온($50-70^{\circ}C$)에서 단시간(2 hr) 동안 열처리한 경우에 trehalose가 함유되지 않은 AYS에 비해 그 활성이 비교적 잘 유지되었다. 따라서 trehalose를 AYS에 첨가하면 열처리 후에도 AYS의 항효모 활성을 다소 보호할 수 있다. Rahnella aquatilis strain AY2000 produces an anti-yeast substance (AYS), however activity of the AYS tends to be decreased by heat. To investigate whether trehalose can protect AYS activity against heat, comparative studies on the AYS with and without trehalose were performed. After heat treatment at high temperatures ($50^{\circ}C-70^{\circ}C$), the AYS with trehalose had higher activity than the AYS without trehalose had. In case of AYS with trehalose (0.3-0.9M), activity of the AYS could be determined at ranging from $7.8\;{\mu}g/mL$ to $31.3\;{\mu}g/mL$ on S. cerevisiae by MTT method. Consequently, activity of the AYS after heating was well maintained by trehalose.

Heterologous expression of bacterial trehalose biosynthetic genes enhances trehalose accumulation in potato plants without adverse growth effects

심재성,서주석,서준성,김용환,구연종,최양도,정춘균 한국식물생명공학회 2019 Plant biotechnology reports Vol.13 No.4

Trehalose is a non-reducing disaccharide found in various organisms, including bacteria, fungi, invertebrates, and plants. It plays essential roles in diverse biological processes such as stress resistance, metabolism, and development in plants. Despite the important role of trehalose, it is challenging to increase trehalose levels because of its negative effects on plant growth and development. In this study, we tested several combinations of different promoters and trehalose biosynthetic genes to efficiently accumulate trehalose in potato plants. Transgenic potato plants expressing TPSP (trehalose-6-phosphate synthase/ phosphatase) fusion gene under the control of Rd29A promoter increased trehalose content in response to high salt stress and did not show significant growth retardation. Although constitutive overexpression of plastid-targeted MTSH (maltooligosyltrehalose synthase/trehalohydrolase) fusion gene driven by CaMV 35S promoter remarkably accumulated trehalose, the transgenic potato plants showed a severely stunted phenotype. By contrast, stress-inducible expression of plastid-targeted MTSH under the control of Rd29A promoter led to trehalose accumulation without growth defects in response to high salt stress. Besides, tuber-specific expression of plastid-targeted MTSH driven by GBSS (granule-bound starch synthase) promoter specifically increased trehalose level in potato tubers without stunting growth. Consequently, we suggest that heterologous expression of TPSP and MTSH fusion genes driven by stress-inducible or tuber-specific promoters can be an effective strategy for accumulating trehalose in potato plants as well as minimizing its adverse effects on potato growth.

Trehalose에 의하여 동결 건조된 정자의 돼지 난자 내 직접주입 후 체외 배발달

강화형,이지웅,강만종,김광현,문승주 韓國受精卵移植學會 2014 한국동물생명공학회지 Vol.29 No.1

본 연구는 돼지 정자의 동결 건조 시 동결 보호제인 trehalose의 효과와 동결 건조 시간과 동결 건조 후 저장 기간에 따라 정자의 생존성과 체외 성숙 난자 내 동결 건조 정자를 직접 주입한 후 전핵 형성율, 난할율 그리고 배발달 성적을 조사하였다. 동결 건조 후 정자의 생존율은 trehalose 무첨가구에 비해 trehalose를 첨가한 처리구에서 높은 생존율을 보였으며, 75 mM의 trehalose를 첨가하여 동결 건조한 정자들의 생존율이 가장 높은 것으로 나타났다. 또한, 동결 건조 후 저장 기간이 길어질수록 생존율이 낮아지는 경향이었다. 체외 성숙 난자 내 동결 건조 정자를 직접 주입 후 전핵 형성율은 trehalose 첨가구에서 유의적으로 높았으며(p<0.05), 난할율과 배발달 성적도 trehalose 첨가구에서 유의적으로 높았고(p<0.05), 정자의 동결 건조 시간이 짧을수록 높은 난할율과 배발달율을 보였다. The objectives of this study were to investigate the effects of trehalose as a cryoprotectant for porcine freeze-dried spermatozoa, to find the optimal freeze-drying time and storage periods of freeze-dried spermatozoa, and to find out pronuclear formation rates, cleaved rates, and embryo development through intracytoplasmic injection of freeze-dried spermatozoa on porcine oocytes. The survival rates of spermatozoa after freeze-drying with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). The highest survival rates were found at 75 mM trehalose treatment. The longer storage periods after freeze-drying seemed to have a lower survival rates. Development in culture of pig by ICSI with trehalose treatment were significantly higher than those of them without trehalose treatment (p<0.05). Shorter freeze-drying time of spermatozoa was resulted in the highest cleaved rates and embryo development.

Trehalose가 식빵의 품질 특성에 미치는 영향

김영자(Young-Ja Kim),이정훈(Jeong-Hoon Lee),정구춘(Koo-Chun Chung),이시경(Si-Kyung Lee) 한국식품영양과학회 2014 한국식품영양과학회지 Vol.43 No.5

Trehalose가 식빵의 품질 특성에 미치는 영향을 평가하기 위하여 밀가루 대비 설탕을 6% 첨가한 것을 대조구로 하여 설탕 6%와 함께 trehalose를 각각 2, 4, 6% 첨가하여 식빵을 제조하였다. 부피 및 비용적, 굽기손실률, 조직감, 수분 함량, 수분활성도, crumb 색도 등을 분석하였고 관능검사를 실시하였다. 부피와 비용적은 trehalose 4% 첨가구가 2,140 mL 및 3.96 mL/g으로 가장 컸고, 굽기손실률은 6% 첨가구가 9.07%로 가장 적었다. 조직감 분석에서 경도는 저장 7일까지 trehalose 4% 첨가구가 가장 낮았다. 수분 함량과 수분활성도는 저장 7일 동안 trehalose 6% 첨가구가 가장 높았고, crumb 색도는 6% 첨가구가 가장 밝은 것으로 나타났다. 관능검사에서 trehalose 4% 첨가구가 96.1점으로 가장 높은 점수를 얻었다. 이상의 trehalose 첨가량을 달리한 식빵 제조 실험에서 제품 특성에 긍정적인 영향을 주는 것으로 나타나 빵에 활용 시 품질 향상에 효과가 있을 것으로 사료되었다. This study was carried out to evaluate the effects of trehalose on quality characteristics of white pan bread. Basic formula consisted of bread flour, 6% sucrose, and 0, 2, 4, 6% trehalose, respectively. Loaf volume, specific loaf volume, baking loss rate, moisture content, water activity, crumb color, and sensory evaluation scores were determined for quality characteristics of bread. White pan bread containing 4% trehalose had a higher loaf volume of 2,140 mL and specific loaf volume of 3.96 mL/g. Baking loss rate of white pan bread was the lowest in the loaf containing 6% trehalose. In crumb texture analysis, hardness of loaf containing 4% of trehalose was the lowest after 7 days of storage. Moisture content and water activity of white pan breads increased with increasing level of trehalose. Color of crumbs was the brightest in the loaf containing 6% trehalose. White pan bread containing 4% trehalose scored better in the sensory evaluation than the others. As a result, trehalose affected quality characteristics of white pan bread. Especially, the addition of 4% trehalose to white pan bread had beneficial effects on quality characteristics.

형광성 Pseudomonas, SSL3 균주의 생존율에 미치는 Trehalose의 효과

성기영,류옥란,최원열,Seong, Ki-Young,Ryu, Ok-Ran,Choi, Won-Yeol 한국토양비료학회 2000 한국토양비료학회지 Vol.33 No.4

유용한 기능을 갖는 미생물을 농업현장에 쉽게 이용할 수 있게 하기 위하여 일반적으로 이들 균을 담체와 함께 건조시켜 입단화나 분말화 시킨다. 생체균을 건조하는 과정에서 미생물은 수분손실로 인해 삼투현상에 직면하며 거의 대부분이 사멸하고 만다. 본 연구에서는 높은 염농도나 건조에 의해 야기되는 균의 사멸을 감소시키기 위해 삼투보호제로 알려진 trigonelline이나 trehalose가 균의 생존에 미치는 영향을 조사하였다. 형광성 Pseudomonas, SSL3 균주의 염내성 농도는 5%가 한계농도이며, 7% 이상에서는 생장이 불가능하였다. 염농도가 높은 액체 배지에 trigonelline을 첨가하여 균의 생육을 조사한 결과 trigonelline은 배지의 종류나 염농도와 상관없이 균의 생장을 억제하는 효과가 있었다. 한편 trehalose를 액체배지에 첨가하면 10mM 농도이하에서는 4% NaCl을 포함한 LB 액체배지에서도 균의 생장이 증진되었으며 150mM이상의 농도에서는 균의 생장이 억제되었다. 균을 건조분말화시킬 때 trehalose를 첨가하여 토양중에서 균의 생존율을 조사한 결과 건조토양에서도 액체배지에서와 마찬가지로 trehalose를 첨가하면 첨가하지 않은 대조구에 비하여 균의 생존율이 증진되었다. Trehalose를 첨가한 제재화 균은 수분이 충분한 토양이나 건조한 토양 어느쪽에서든지 대조구에 비하여 균의 생존이 증진되었으며 특히 미생물 제제를 $-20^{\circ}C$나 $-70^{\circ}C$에 보관한 토양의 경우에 trehalose가 첨가된 제제의 생존율이 월등히 뛰어났다. FTIR의 분석결과 trehalose를 첨가한 4% 염농도에서 생장한 균에서는 대조구에 비하여 2차 amide기의 변화가 경감되었다. 이 결과에서 보듯이 trehalose는 염농도나 한발조건에 의해서 야기되는 균의 수분 스트레스로부터 균의 외막을 보호하여, 단백질구조의 급격한 변화를 감소시키므로 균의 생존율이 증진되는 것으로 사료된다. To make a better use of the beneficial bacterial inoculants in the agricultural practice, dry forms of bacterial fertilizer or pesticides are prepared with carrier materials. During the drying process of bacterial inoculant, most of the cells face a severe osmotic pressure and dehydration, and die off. Our study describes the effect of osmoprotectants such as trigonelline and trehalose on the survival of bacterial cells in high salt concentration and drying conditions. A fluorescent Pseudomonas, strain SSL3, used in this study, could grow in high salt concentration of upto 5% but the cells could not overcome the growth retardation at over 7% of salt concentration. The addition of trigonelline, even on small amount, in liquid medium containing 4% NaCl was detrimental to the cell. However, the addition of trehalose of upto 10 mM to the liquid medium containing 4% NaCl, enhanced cell growth. The cell growth was retarded when 150mM trehalose was added to the medium. Upon dry formulation of cells, trehalose was added. And the dry cells were inoculated into the soil to determine the effect of osmoprotectants on the survival of the cells. The survival of the cells, both in wet or dry soil, was improved by the addition of trehalose during the dry cell formulation. The positive effect of trehalose on the cell survival at $-20^{\circ}C$ and $-70^{\circ}C$ was oven more pronounced. The FTIR (Fourier transformation infra-red) spectroscopic analysis showed that the change of the 2nd amide group was reduced by adding trehalose to the medium containing 4% NaCl. These results suggest that trehalose can protect the cell membrane from dryness or high concentration of salt, thereby diminishing the sudden change of the protein structure of the cell membrane and, as a consequence, improving the cell survival.

Molecular characterization and functional analysis of two trehalose transporter genes in the cabbage beetle, Colaphellus bowringi

Li Jia-Xu,Cao Zhen,Guo Shuang,Tian Zhong,Liu Wen,Zhu Fen,Wang Xiao-Ping 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.3

The trehalose, major blood sugar in insects, enhances stress resistance of diapausing individuals in adverse environment and provides an energy source for reproduction. Trehalose transporters (TRETs) play an important role in transport of trehalose from trehalose-producing tissues, e.g. fat body, to trehalose-consuming tissues. Although studies have shown that trehalose contributes to diapause and reproduction, the function of TRETs in these processes remains unclear. In this work, we cloned two TRET genes, TRET1a and TRET1b, from the cabbage beetle Colaphellus bowringi, which is capable of entering reproductive diapause under long-day conditions. We also analyzed the expression profiles of these two genes and investigated their potential roles in diapause and reproduction. The results suggested that both TRET1a and TRET1b belong to sugar-transporter and major facilitator superfamilies. Interestingly, TRET1a was highly expressed in the fat bodies of diapause-destined (DD) females but TRET1b was predominantly expressed in the ovaries of non-diapause-destined (NDD) females. Hormonal induction indicated that juvenile hormone induced TRET1b but repressed TRET1a at transcriptional levels. Methoprene-tolerant and Krüppel homolog 1 mediated the JH-suppressed TRET1a expression but were not involved in the regulation of TRET1b expression by JH. RNAi of TRET1a in DD females elevated the trehalose content in the fat bodies and suppressed the expression of a couple of genes related to stress resistance, which is a critical diapause trait. Knockdown of TRET1b in NDD females reduced the trehalose content in the ovaries but had no apparent effect on the ovary development and yolk deposition. These data suggest that TRET1a and TRET1b could regulate the trehalose content in specific tissues and may play potential roles in reproductive diapause in the females of C. bowringi. Introduction Diapause is a survival strategy for many insects in adapting to adverse environmental conditions (Tauber and Tauber, 1976; Tougeron, 2019). Diapause is a dynamic successive process that consists of diapause induction, preparation, initiation, maintenance, termination, and post-diapause quiescence (Kostal, 2006). Insects accumulate enormous nutrients such as sugars and lipids to enter diapause during diapause preparation phase (Hahn and Denlinger, 2007, 2011). The trehalose is the major hemolymph sugar in insects. The sugar not only provides source of energy but also protects proteins and cellular membranes from dehydration, desiccation, heat, cold, and oxidation (Elbein et al., 2003; Li et al., 2002; Richards et al., 2002; Wyatt and Kalf, 1957; Yancey, 2005; Zhu et al., 2008). Therefore, trehalose can play roles in reproduction and diapause by mediating energy supply and stress tolerance, respectively (Kamei et al., 2011; Lu et al., 2019; Xu et al., 2009).

계육 Surimi에 Trehalose와 Oligosaccharide의 냉동변성 방지효과

이성기,민병진 한국가금학회 2002 韓國家禽學會誌 Vol.29 No.1

계육 surimi에서 trehalose와 oligosaccharide의 단백질 냉동변성 방지효과를 구명하기 위하여 본 실험을 실시하였다. 기계발골노계육에 0.5% 소금물(고기:용액, 1:4)로 2회, 증류수로 1회 수세하여 surimi를 제조하였다. Trehalose와 oligosaccharide를 각각 8%씩 혼합한 후 $-18^{\circ}C$에서 10주간 냉동저장하였다. 모든 Surimi의 적색도(a*)는 냉동기간동안 감소하였다. 냉동기간중에 trehalose, oligosaccharide, 무첨가 순으로 명도가 높고 적색도와 황색도가 낮아 색택 안정성이 있는 것으로 나타났다. 냉동기간중 surimi의 gel 강도(압착력, 경도, 탄성, 점착성)는 감소하였지만, trehalose와 oligosaccharide 첨가가 무침가구에 비해 유의적으로 높은 수준을 유지하였다. 냉동기간중에 부착성은 trehalose 첨가가 가장 높았다. 따라서 이들 당류가 계육 surimi에 단백질 냉동변성방지제로 효과가 인정되었으며, trehalose는 감미도가 기존 설탕보다 낮고 색깔 및 물성에서 oligosaccharide 보다 더 우수한 것으로 판단되었다. Cryoprotective effects on chicken surimi during storage were investigated. Chicken surimi from mechanically deboned spent layer meat was prepared with 4 volumes of 0.5% NaCl washing, and then blended with or without cryoprotectants (8% trehalose, 8% oligosaccharide) prior to frozen storage at $-18^{\circ}C$ to 10 weeks Redness (a) of all surimi decreased during storage. Color stability increased during storage when lightness increased but redness decreased. At this Point, surimi maintained a better color quality as followed order; trehalose > oligosaccharide ) non-additive. Gel strength such as compressive force, hardness, adhesiveness and gumminess tended to decrease during frozen storage. Cryoprotectants provided significantly better textural properties than non-auditive. Surimi with trehalose showed the highest adhesiveness. In conclusion, trehalose and oligosaccharide seemed to be good cryoprotectants of chicken surimi. Especially, trehalose resulted in better cryoprotectant than oligosaccharide because of better color stability, better textural properties, and lower sweet characteristics.

Cryopreservation with Trehalose Reduced Sperm Chromatin Damage in Miniature Pig

박철호,김성원,황유진,김대영 사단법인 한국동물생명공학회 2012 한국동물생명공학회지 Vol.27 No.2

Miniature pig sperm cryopreservation is continually researched in biotechnology for breed conservation and reproduction. It is important to control the temperature at each stage of cryopreservation and cryoprotectant. It is also necessary to find the optimal cryoprotectant concentration and chemical elements of the extender. Recently, many studies have used various cryoprotectant materials, such as dimethyl sulphoxide (DMSO), ethylene glycol (EG), antifreeze protein (AFP), amides, and glycerol. Glycerol is a commonly used cryoprotectant. However, glycerol has critical cytotoxic properties, including osmotic pressure and it can cause irreversible damage to live cells. Therefore, We focused on membrane fluidity modifications can reduce cell damage from freezing and thawing procedures and evaluated on the positive effects of trehalose to the viability, chromatin integrity, and motility of boar sperm. Miniature pig sperm was separated from semen by washing with modified- Modena B (mMB) extender. After centrifugation, the pellet was diluted with the prepared first extender. This experiment was designed to compare the effects that sperm cryopreservation using two different extenders has on sperm chromatin. The control group used the glycerol only and it was compared with the glycerol and glycerol plus trehalose extender. Sperm viability and motility were evaluated using WST1assays and computer-assisted semen assays (CASA). Chromatin structure was examined using acridine orange staining. For the motility descriptors, trehalose caused a significant (p<0.01) increase in total motility (57.80 ± 4.60% in glycerol vs. 75.50 ± 6.14% in glycerol + trehalose) and progressive (51.20 ± 5.45% in glycerol vs. 70.74 ± 8.06% in glycerol +trehalose). A significant (p<0.05) increase in VAP (42.70 ± 5.73 μm/s vs. 59.65 ± 9.47 μm/s), VSL (23.06 ± 3.27μm/s vs. 34.60 ± 6.58μm/s), VCL (75.36 ± 11.36 μm/s vs. 99.55 ± 12.91μm/s), STR (54.4 ± 2.19% vs. 58.0 ± 1.63%), and LIN (32.2 ± 2.05% vs. 36.0 ± 2.45%) were also detected, respectively. The sperm DNA fragmentation index was 48.8%to glycerol only and 30.6% to glycerol plus trehalose. Trehalose added group showed higher percentages of sperm motility,stability of chromatin structure than glycerol only. In this study, we suggest that trehalose is effective in reducing freezing damage to miniature pig sperm and can reduce chromatin damage during cryopreservation.