Natural environmental resources are considered a prospective source of microorganisms capable of producing biocatalysts with great potential in industrial areas. Arable soil fertilized with peat moss is a habitat for various microorganisms. The present research focused on the isolation and identification of hydrolase-producing bacteria that thrive at a broad temperature range. In this study, a total of 33 strains were isolated from arable soil fertilized with peat moss (Silla Garden in Busan, South Korea). The isolated bacteria were mesophiles and thermophiles with a wide temperature range. Taxonomic identification showed that the isolated strains belonged to 2 phyla, 5 families, 10 genera, and 24 species. Subsequently, the isolated strains were screened for hydrolase (amylase, lipase, and protease) activity. All isolates possessed activity of at least one enzyme and six bacterial isolates produced combined extracellular enzymes. Diversity of soil bacteria species in the present study suggest the potential of soil bacteria in the various industrial applications.

In order to isolate thermophilic bacteria with high activity of CMCase and xylanase, spent mushroom substrates was collected from an oyster mushroom cultivation farm in Jinju, Gyeongnam, Korea. Among the isolates, one strain designated as YJ09 was selected by agar diffusion method. The isolate YJ09 was identified as a member of Bacillus licheniformis based on biochemical characteristics using Bacillus ID kit and MicroLog system. Comparative 16S rDNA sequence analysis showed that isolate YJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus licheniformis with sequence similarity of 98.9%. Based on its physiological properties, biochemical characteristics and phylogenetic distinctiveness, the isolate YJ09 was classified as Bacillus licheniformis. The CMCase and xylanase activity of B. licheniformis YJ09 was slightly increased corresponding to the bacterial population from exponential phase to stationary phase in the growth curve of B. licheniformis YJ09.

We have used bulked segregant analysis to screen the strain-specific DNA marker associated thermophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 550 bp was amplified with OP-S07 primer from the thermophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-S07-1. The PCR analysis with the OP-S07-1 primer showed that this SCAR marker clearly distinguish the thermophilic strains from the control strains.

We have used bulked segregant analysis to screen the strain-specific DNA marker associated thermophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 500 bp was amplified with OP-A11 primer from the psychrophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-A11-1. The PCR analysis with the OP-A11-1 primer showed that this SCAR marker clearly distinguish the psychrophilic strains from the control strains.

음식물 폐기물 침출수를 처리하는 분리막 결합 고온 혐기성소화공정(생물학적 반응조) (Anaeorobic Membrane Bioreactor, AnMBR)의 파일럿 운전에서 분리막의 교차여과 속도와 막간압력이 파울링에 미치는 영향을 관찰하였다. 연구 결과 정압여과 하에서 교차여과 속도가 증가할수록 파울링의 속도는 현격히 감소되었다. 그러나 이와 같은 영향은 낮은 막간압력에서 더욱 효과적이었다. 막간압력이 증가할수록 여과대상 물질의 압축성으로 인해 투과성이 상대적으로 낮은 파울링층(혹은 케익층)이 분리막 표면에 형성된 것에 기인된 듯하다. 여과대상 시료의 입도분석을 해 본 결과 입자크기는 약 10~100mum 범위에서 분포하였고 이에 따라 브라운확산에 의한 역수송보다 분리막 표면에서 교차여과에 의해 발생하는 전단력이 입자의 역수송에 더욱 기여하고 있음을 예측할 수 있었으며 이는 AnMBR의 연속운전을 통해 재확인할 수 있었다. 운전 후 막 부검을 실시한 결과 유기 및 무기 파울링이 모두 관찰되었으나 어느 것이 지배적인 파울링 기작을 나타내는지는 앞으로 더욱 연구가 필요하다. 무기 파울링의 경우 대부분 분리막 표면에서 스케일링 형성이 지배적이었으며, 따라서 분리막의 공극 막힘에 주로 기여하는 작은 콜로이드성 유기물질의 경우 분리막 표면에서 전단력에 의한 역수송 효과는 그다지 크지 않을 것으로 사료된다.

In order to isolate thermophilic compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated HJ01 was selected by agar diffusion method. The strain HJ01 was identified as members of the Bacillus subtilis by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain HJ01 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rDNA gene sequence similarity of 99.3%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain HJ01 was classified within the genus Bacillus, for which the name Bacillus subtilis HJ0 is proposed. The cellulase and xylanase activity of B. subtilis HJ0 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for B. subtilis HJ01.

Spent mushroom substrate(SMS) is a by-product remaining after a crop of mushrooms. About 9 thermophilic strains were isolated from SMS(Flammulina velvtipes). Among of them, one isolate, designated UJ09, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain UJ09 was produced cellulase and xylanase as hydrolase. The strain UJ21 was identified as members of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain UJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with 16S rDNA gene sequence similarity of 99.2%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain UJ09 was classified within the genus Bacillus, for which the name Bacillus amyloliquefaciens UJ09 is proposed.

호열성 미생물을 검토하기 위하여 전국 각지로부터 토양과 두엄을 채취하여 그로부터 호열성 미생물을 분리하였다. 토양과 두엄으로부터 분리된 호열성 미생물 1250여 균주를 선별하였고, 이들을 대상으로 미생물이 생산하는 효소활성을 검토하여 호열성 전분 분해 효소를 생산하는 1주의 미생물을 확인하였다. 확인된 1주의 미생물을 strain 2719라 명명하였다. Strain 2719 균주는 형태학적으로 gram 양성 간균의 특징을 나타냈고, 균주의 표면은 매끄럽지 않았으며, 비교적 다양한 길이를 가지고 있었다. 또한 다른 gram 양성 간균들에 비해서 많은 수의 균사들이 각 균주들 사이에 복잡하게 얽혀있었다. 생화학적 특성을 확인한 결과 catalase 양성, glucose 발효, arabinose 발효, mannitol 발효, casein·gelatin·starch 가수분해의 특징을 가지고 있었으며, 이는 Bacillus sp.로 추정되었다. 생육 pH의 범위는 pH 6-pH 8 범위에서 생육이 가능했으며, 생육 온도의 범위는 50-70oC였다. 16S rDNA sequence 분석결과 Bacillus thermoglucosidasius의 16S rDNA와 99.52%가 일치하였으나, sequence의 일부분이 다른 부분이 있고, 생육 특성에서 약간의 차이를 보였다. 또한 gene bank에 등록되어 있는 균주들의 16S rDNA sequence들과 비교하여도 일치하는 균주는 확인되지 않았다. 이와 같은 실험결과에 따라 2719 균주는 기존에 발표되지는 않았으나, Bacillus thermoglucosidasius와 매우 유사한 균주로 판단되어 Bacillus thermoglucosidasius 2719로 명명하였다.

This study was performed to test the feasibility of thermophilic (55oC) co-digestion of municipal sewage sludge and food wastes. The management variables of co-digestion were the mixed ratios of municipal sewage sludge and food waste hydraulic retention times (HRTs). During the operation of thermophilic co-digestion, the reactor pH ranged from 7.0 to 7.5 and the reactor alkalinity remained above 3,200 to 4,000 mg/L as CaCO3. The volatile fatty acids concentration increased as the HRT shortened from 20 days to 10 days and the mixture ratio increased to 1:4, but did not reach toxic levels for co-digestion of sewage sludge and food wastes. Methane productivity increased gradually as the organic loading rate increased. Maximum methane productivity reached 1.03v/v-d at an HRT of 10 days and at the mixture ratio of 1:4. The TVS removal efficiency decreased from 70.6% to 58.3% as the HRT shortened from 20 days to 10 days. TVS removal efficiency ranged from 57.0% to 77% during the entire operation. It is likely that thermophilic co-digestion of sewage sludge and food wastes is a very effective method both to environmentally treat food waste and to economically produce gas for energy.

하수처리과정에서 발생하는 슬러지의 부피를 줄이는 동시에 이들의 유기물 성분을 메탄 등의 바이오가스로 전환하기 위해 중온(35℃)에서의 혐기소화 공정이 널리 적용되고 있다. 혐기소화공정의 안정성이나 에너지 투입량 측면에서는 중온혐기소화가 적합하다고 알려져 있지만, 높은 유기물 부하량(organic loading rate, OLR)을 처리하기 위해 반응속도를 상승시키거나 SRT(sludge retention time)을 줄이기 위해 고온(55℃) 혐기소화를 적용하기도 한다. 고온 혐기소화공정을 새롭게 시작할 때 접종물을 기존의 고온 혐기소화공정으로부터 얻기 힘든 경우 중온혐기소화액을 고온에서 적응시켜 접종하기도 한다. 이때 온도를 적응시키는 방법에 따라 공정 효율이 달라지는데, 연구에서는 중온혐기소화액으로부터 고온 혐기소화 접종물을 제작하기 위한 방법으로 온도를 올리는 방법을 달리하였을 때 이들의 메탄 생산과 미생물 군집에 미치는 영향을 살펴보고자 하였다. 본 연구수행을 위해 광주 제 1 하수처리장에서 중온혐기소화액, 1, 2차 농축 슬러지 등을 샘플링 하여 두 대의 반응기(Working volume : 3L)를 설치하여 회분식(Batch)운전 후 연속교반탱크형반응기(CSTR)로 운전(SRT 20, 40일)하였다. 먼저 한 대의 반응기는 35℃에서 55℃로 한 번에 온도변화를 주었고, 다른 반응기의 경우에는 35℃에서 하루에 1도씩 올려서 약 20일에 걸쳐 55℃로 올린 뒤 55℃로 유지하였다. 반응기의 효율을 확인하기 위해 메탄 발생량, total solids (TS), volatile solids (VS), total chemical oxygen demand (tCOD), soluble chemical oxygen demand (sCOD), soluble components (protein, carbohydrate), pH 등을 측정하였으며, NGS (next generation sequencing)를 활용하여 혐기소화 전/후의 소화액의 미생물 군집변화를 알아보았다.

혐기성소화는 폐기물처리뿐만 아니라 대체에너지인 메탄가스가 발생하여 에너지를 회수할 수 있다는 장점을 가지고 있다. 혐기성소화 효율을 높이기 위해 미량중금속을 투입하기도 한다. 이는 대표적인 유기성 폐기물인 음식물류폐기물에 미량중금속의 함량이 낮고, 미량중금속이 미생물의 생장에 중요한 역할을 하기 때문이다. 이에 본 연구에서는 미량중금속의 함량이 낮고 대표적인 유기성폐기물인 음식물류폐기물을 기질로 사용하여 혐기성소화를 진행하였고 식종은 음식물류폐기물과 축산폐수를 병합처리하는 혐기성소화조 소화슬러지를 사용하였다. 사용한 시료는 A시의 폐기물자원화시설에서 채취하였으며 음식물류폐기물은 2 mm, 소화슬러지는 0.85 mm 채로 걸러서 사용하였다. Ni는 메탄생성미생물의 성장에 필수적으로 필요하고 아세트산의 이용효율을 증가시키는 효과가 있어 대상 미량중금속을 Ni로 하였다. Ni 투입량은 0, 0.1, 1, 10, 50 mg/L로 변화를 주었으며 소화온도는 중온(35℃)과 고온(55℃)로 실험을 진행하였다. 실험은 음식물류폐기물과 소화슬러지를 휘발성고형물(volatile solid, VS) 기준으로 1:1 비율로 섞어 500 mL serum bottle에 300 mL를 채워 진행하였고, 반응기에 농도별로 Ni를 투입하였다. 이 후 질소를 이용하여 2분간 퍼지하고, 고무마개와 알루미늄씰(aluminium seal)을 이용하여 밀봉하였다. 제작된 반응기를 항온 진탕배양기(VS-8480, VS-8480SF, KR)에서 중온 및 고온에서 소화를 진행하였다. 발생하는 가스는 기압계(Keller LEO-2, Germany)를 이용하여 반응기 내 압력을 측정하여 발생량을 계산하는 방식을 이용하였다. 본 연구는 아직 진행 중에 있어 결과는 추후 학회에서 발표할 예정이다.

This study was conducted to evaluate the effects of physical characteristics. Twelve specific odorous compounds and various sources of bacteria were tested via treatment of food waste using an ultra-thermophilic aerobic composting process. Food waste was mixed with seed material and operated for 47 days. During composting, the temperature was maintained at 80-90°C. The variations in O2, CO2 and NH3 production suggested typical microorganism-driven organic decomposition patterns. After composting, the concentrations of 12 specific odorous compounds other than ammonia did not exceed the allowable exhaust limits for odor. After composting, thermophiles represented 50% of all bacteria. After composting, the percentage of thermophile bacterial increased by 15%. Therefore, both stable composting operation and economic benefit can be expected when an ultra-thermophilic composting process is applied to food waste.

Batch cultivations were performed to evaluate the influences of the initial pH condition on mesophilic and thermophilic acidogenic fermentation with food waste recycling wastewater. In both conditions of mesophilic and thermophilic fermentation, TVFAs production rates were maximized at the initial pH 7 condition as 0.15 and 0.23 g TVFAs/L·hr, respectively. And pH was also maintained stably between 6 and 7 during 72hr acidogenic cultivation at both conditions. However, predominant VFA components were different according to reaction temperature conditions. In mesophilic condition, propionic acid which has low conversion efficiency to methane was accumulated up to 1,348 mg/L while acetic and butyric acid were predominant in thermophilic condition. Therefore, thermophilic acidogenic fermentation was superior for the effective VFAs production than mesophilic condition. From the DGGE analysis, the band patterns were different according to the initial pH conditions but the correlations of the each band were increased in similar pH conditions. These results mean that microbial communities were certainly affected by the initial pH condition. Consequently, the adjustment of the initial pH to neutral region and thermophilic operation are needed to enhance acidogenic fermentation of food waste recycling wastewater.