KR20220053683A - 단일 세포 단백질 또는 바이오매스 생산을 위한 균주 및 방법 - Google Patents
단일 세포 단백질 또는 바이오매스 생산을 위한 균주 및 방법 Download PDFInfo
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Abstract
본 발명은 크산토박터 속의 박테리아 균주 및 크산토박터 속의 박테리아를 사용하여 단백질 또는 바이오매스를 생산하기 위한 연속 배양 방법에 관한 것으로서, 상기 방법은 기체 및 미네랄을 세포에 공급하는 단계를 포함한다. 본 발명은 또한 이러한 방법의 산물 및 이러한 산물의 예를 들어 식품 또는 사료에서의 용도에 관한 것이다.
Description
본 발명은 미생물을 이용한 단백질 및/또는 기타 거대분자의 생산에 관한 것이다. 구체적으로, 본 발명은 신규한 박테리아 균주, 및 박테리아를 이용하여 단백질 또는 바이오매스를 생산하기 위한 연속 배양 방법에 관한 것으로서, 여기서 기체 및 미네랄이 세포에 공급된다. 본 발명은 또한 이러한 방법의 산물, 및 이러한 산물의 예를 들어 식품 또는 사료에서의 용도에 관한 것이다.
세계 인구 증가, 기후 변화 및 물 부족은 전통적인 농업 및 이에 따른 식량 및 사료의 충분한 공급을 점점 더 위협하고 있다. 그러므로, 유기 분자 예컨대 단백질의 대체 공급원을 조사하였다. 잠재적인 대안으로는 단일 세포 생산, 즉 미생물을 이용하여 단백질 및/또는 기타 거대분자를 생산하는 것이다.
화학독립영양 미생물 (chemoautotrophic microorganisms)은 에너지 공급원으로서 수소 기체 및 유일한 탄소 공급원으로서 이산화탄소를 사용하여 최소 미네랄 배지 (minimal mineral medium)에서 성장할 수 있다고 개시되어 있다. 이러한 미생물에 대한 검토는 예를 들어 Shively et al. (1998) Annu Rev Microbiol 52:191를 참조한다. 특허출원 WO2018144965는 기체 기질을 고-단백질 바이오매스로 전환하기 위한 다양한 미생물 및 생물공정을 개시하였다. Andersen et al. (1979) Biochim Biophys Acta 585:1-11은 종속영양 (heterotrophic) 및 자가영양 (autotrophic) 조건에서 쉽게 성장하는 수소 박테리아인 알칼리게네스 유트로푸스 (Alcaligenes eutrophus)의 돌연변이 균주를 개시하였다. 리불로스-1,5-비스포스페이트 카복실라제/옥시게나제 (rubisco) 활성이 변경된 돌연변이체를 특성규명하였다. Ohmiya et al. (2003) J. Biosci. Bioeng. 95:549-561은 취급이 어려운 (recalcitrant) 바이오매스 활용에 대한 미생물 유전자의 적용을 검토하였다. Yu Jian et al. (2013) Int J Hydrogen Ener 38:8683-8690은 수소-산화 박테리아 단리물에 의한 이산화탄소 고정을 개시하였다. 50%의 높은 에너지 효율이 적당한 산소 농도 (10 mol%) 하에 측정되었다.
그러나, 다양한 화학독립영양 미생물은 성장 속도, 수율, 바이오매스 조성뿐만 아니라 식품 성분으로서의 사용과 관련된 특성 예컨대 인간 섭취의 안전성, 맛, 냄새, 마우스필 (mouth-feel), 조리시 기술적 및 기능적 특성 등의 측면에서 서로 다른 특성을 가지고 있다. 모든 화학독립영양 미생물이 충분한 성장 속도 및 충분한 수율을 제공하는 것은 아니며, 모든 공정이 경제적으로 실행 가능한 대규모 공정으로 현실적으로 확장될 수 있는 것은 아니다. 예를 들어 식품 또는 사료 적용을 위해 기능성 단백질을 충분히 생산하기 위해서는 대규모로 수행될 수 있는 적절한 공정 및 적절한 생산 유기체를 찾는 것이 중요하다. 이러한 필요성이 본 발명에 의해 다루어진다.
발명의 요약
제1 주요 양상에서, 본 발명은 단리된 박테리아 균주 VTT-E-193585 또는 이의 유도체에 관한 것이다.
추가 양상에서, 본 발명은 본 발명의 박테리아 균주 또는 이의 유도체를 포함하는 배양물에 관한 것이다. 또한, 본 발명은 바이오매스 및/또는 단백질의 생산 방법에 관한 것으로서, 상기 방법은 본 발명의 박테리아 균주 또는 이의 유도체를 배양하는 단계를 포함한다.
추가 양상에서, 본 발명은 바이오매스 및/또는 단백질의 생산 방법에 관한 것으로서, 상기 방법은 에너지 공급원으로서 수소 및 무기 탄소 공급원을 사용하여 연속 배양으로 크산토박터 (Xanthobacter) 속의 박테리아 균주를 배양하는 단계를 포함하고, 상기 무기 탄소 공급원은 이산화탄소를 포함한다.
추가의 주요 양상에서, 본 발명은 본 발명의 방법에 의해 수득되거나 또는 수득 가능한 벌크 단백질, 바이오매스 또는 비-단백질 세포 또는 화학 성분, 및 본 발명의 방법에 의해 수득되거나 또는 수득 가능한 식품 또는 사료 제품에 관한 것이다.
도 1. VTT-E-193585로 기탁된 단리된 박테리아 균주의 화학독립영양 200-L 배양 중에 600 nm에서 측정된 광학 밀도 (검정색 원) 및 광학 밀도 프로브 판독값.
도 2. 상이한 질소 공급원들에 대한 VTT-E-193585로 기탁된 단리된 박테리아 균주의 병렬 화학독립영양 200-mL 배양 중에 600 nm에서 측정된 광학 밀도.
도 2. 상이한 질소 공급원들에 대한 VTT-E-193585로 기탁된 단리된 박테리아 균주의 병렬 화학독립영양 200-mL 배양 중에 600 nm에서 측정된 광학 밀도.
발명의 상세한 설명
정의
예를 들어 균주의 맥락에서 용어 "단리된"이 본원에서 사용되는 경우, 이의 자연 환경으로부터 단리되는 것을 의미한다. 바람직하게는, 단리된 균주는 순수하며, 즉 다른 균주는 없다.
균주의 맥락에서 용어 "유도체"가 본원에서 사용되는 경우, 참조 균주로부터 유래되는, 즉 참조 균주를 출발점으로서 사용하여 생성된 균주를 지칭한다. 예를 들어, 유전자-조작되거나 또는 달리 돌연변이되거나 또는 유전자-변형된 균주는 이러한 유도체의 구체예이다. 유전자 변형은 전체 유전자좌 또는 이의 단편의 삽입 또는 결실을 포함하는 삽입 또는 결실 뿐만 아니라, 점 돌연변이를 포함한다. 유도체는 바람직하게는 참조 균주와 비교하여, 10개 미만의 유전자 변형, 예를 들어 5개 미만, 예컨대 4, 3, 2 또는 1개의 유전자 변형(들)을 갖는다.
명사 "배양물"이 본원에서 사용되는 경우, 액체 배지 중에 생존 세포의 현탁액을 지칭한다.
용어 "바이오매스"는 박테리아 발효 분야에서 이의 통상적 의미를 가지며, 세포 물질을 지칭한다.
용어 "연속 배양"이 본원에서 사용되는 경우, 새로운 배지가 배양에 연속적으로 부가되고, 박테리아 배양물이 있는 배지가 본질적으로 동일한 속도로 연속적으로 제거되는 배양 과정을 지칭한다.
본 발명의 양상 및 구체예
제1 주요 양상에서, 본 발명은 단리된 박테리아 균주 VTT-E-193585 또는 이의 유도체에 관한 것이다.
균주 VTT-E-193585는 핀란드 Naantali의 발트해 해변에서 단리되었다. 상기 유기체는 제한된 산소 조건에서 에너지 공급원으로서 수소 및 탄소 공급원으로서 이산화탄소와 함께, 최소 미네랄 배지를 사용하여 적절한 생물반응기 조건에서 성장시킬 수 있다. 16S 시퀀싱 및 Illumina 메타유전체학 시퀀싱으로, 상기 균주는 크산토박터 속의 구성원일 가능성이 가장 높지만, 알려진 종은 아닌 것으로 밝혀졌다. 상기 박테리아 균주는 건조된 세포 분말의 단백질 함량이 높고, 모든 필수 아미노산을 함유하기 때문에 식품 및 사료 적용에 매우 적합하다. 이는 또한 포화 지방산보다 더 많은 불포화 지방산 및 높은 수준의 B군 비타민을 함유한다. 알레르기 또는 독성을 유발할 수 있는 펩티도글리칸 및 리포폴리사카라이드의 수준은 낮다. 상기 균주에 대한 독성 분석을 수행하였으며, 유전독성 또는 세포독성이 관찰되지 않았다. 또한, 상기 균주는 일반적으로 항생제에 민감하다.
균주 VTT-E-193585 (SoF1)는 부다페스트 조약에 따른 국제 기탁 기관인, 핀란드 FI-02044 VTT, P.O. Box 1000에 주소를 둔, 핀란드 VTT Technical Research Centre의 VTT Culture Collection에 2019년 6월 11일자로 기탁되었다. 상기 균주의 특성 및 균주 배양 방법에 대한 추가 정보는 본원의 실시예에 제공된다.
바람직한 일 구체예에서, 균주가 균주 VTT-E-193585의 유도체인 경우, 상기 유도체는 에너지 공급원으로서 수소 기체 및 유일한 탄소 공급원으로서 이산화탄소를 사용하여 성장하는 능력을 보유하였다.
일 구체예에서, 균주가 균주 VTT-E-193585의 유도체인 경우, 상기 유도체는 서열번호: 1에 제시된 16S 리보솜 RNA, 또는 서열번호: 1과 최대 20개, 예를 들어 1 내지 10개, 예컨대 1 내지 5개의 뉴클레오티드 차이, 예를 들어 서열번호: 1과 1개, 2개 또는 3개의 뉴클레오티드 차이를 갖는 16S 리보솜 RNA를 포함한다.
서열번호: 1. 균주 VTT-E-193585의 16S 리보솜 RNA 서열:
추가 양상에서, 본 발명은 본 발명의 박테리아 균주 또는 이의 유도체를 포함하는 배양물에 관한 것이다. 바람직한 일 구체예에서, 상기 배양물의 부피는 100 mL 이상, 예를 들어 1 L 이상, 예컨대 10 L 이상, 예를 들어 1,000 L 이상, 예컨대 10,000 L 이상, 예를 들어 50,000 L 이상, 예컨대 100,000 L 이상, 예를 들어 200,000 L 이상이다.
추가 양상에서, 본 발명은 바이오매스 및/또는 단백질의 생산 방법에 관한 것으로서, 상기 방법은 본 발명의 박테리아 균주 또는 이의 유도체를 배양하는 단계를 포함한다. 일 구체예에서, 상기 방법은 바이오매스의 생산을 위한 것이다. 다른 구체예에서, 상기 방법은 단백질의 생산을 위한 것이다. 일 구체예에서, 상기 방법은 에너지 공급원으로서 수소 및 무기 탄소 공급원을 사용하여 연속 배양으로 균주를 배양하는 단계를 포함하며, 상기 무기 탄소 공급원은 이산화탄소를 포함한다. 추가 일 구체예에서, 상기 방법은 바이오매스의 생산을 위한 것이며, 에너지 공급원으로서 수소 및 무기 탄소 공급원을 사용하여 연속 배양으로 균주를 배양하는 단계를 포함하며, 상기 무기 탄소 공급원은 이산화탄소를 포함한다. 상기 방법의 다양한 추가 구체예가 하기에 기재되어 있다.
추가의 주요 양상에서, 본 발명은 바이오매스 및/또는 단백질의 생산 방법에 관한 것으로서, 상기 방법은 에너지 공급원으로서 수소 및 무기 탄소 공급원을 사용하여 연속 배양으로 크산토박터 속의 박테리아 균주를 배양하는 단계를 포함하고, 상기 무기 탄소 공급원은 이산화탄소를 포함한다. 일 구체예에서, 상기 방법은 바이오매스의 생산을 위한 것이다. 다른 구체예에서, 상기 방법은 단백질의 생산을 위한 것이다. 상기 방법의 다양한 추가 구체예가 하기에 기재되어 있다.
게놈 서열에 따르면, 번호 VTT-E-193585로 기탁된 균주는 탄소 고정을 위해 Calvin-Benson-Bassham 사이클을 사용할 가능성 있으며, 여기서 이산화탄소 분자는 리불로스 1,5-비스포스페이트의 5-탄소 사슬에 연결되어 글리세레이트 3-포스페이트의 2개의 분자를 형성한다. 이를 통해 균주는 성장에 필요한 다른 모든 유기 분자를 합성할 수 있다. 수소로부터의 에너지는 NAD+-환원 하이드로게나제 및/또는 NiFeSe-하이드로게나제를 통해 세포로 유입된다. 본질적으로 이는 수소 (H2)가 H+로 산화되고 NAD+가 NADH로 환원되는 산화환원 반응이다. ATP에 추가하여, NADH는 생물체 내부의 주요 에너지 운반체 중 하나이다. 대안으로서, 일부 다른 에너지 등가물은 H2를 사용하는 다른 하이드로게나제 효소에 의해 환원된다. Calvin-Benson-Bassham 사이클은 CO2를 고정하기 위해 ATP 및 NADH/NADPH의 형태로 에너지를 필요로 한다. 상기 균주는 산화적 인산화를 통해 ATP를 생성할 수 있고, 이는 막을 가로질러 양성자 구배를 생성하는 4개의 단백질 복합체로 구성된다. 양성자 구배는 주로 NADH로부터의 에너지를 사용하여 생성된다. 상기 양성자 구배는 ATP를 생성하는 ATP 신타제 복합체를 유도한다. 게놈 서열에 따르면, 상기 균주는 박테리아 F-타입 ATP 신타제를 갖는다.
상기 방법이 무기 탄소 공급원으로 균주를 배양하는 단계를 포함하는 것으로 명시되는 경우, 상기 무기 탄소 공급원은 배양에서 주요 탄소 공급원인 것으로 이해되어야 한다. 따라서, 상기 배양물 중에 유기 탄소 공급원이 소량 존재할 수 있지만, 상기 배양의 주요 대사작용 및 성장은 탄소 공급원으로서 무기 탄소 공급원, 바람직하게는 이산화탄소의 이용에 기반한다. 바람직하게는, 배양에 공급되는 유기 탄소 공급원으로서의 탄소의 비율은 공정 중에 배양에 공급된 모든 탄소의 5% 미만, 예컨대 1% 미만, 예를 들어 0.1% 미만이다. 바람직하게는, 유기 탄소 공급원은 공정에 공급되지 않는다.
유사하게, 상기 방법이 에너지 공급원으로서 수소 (H2)를 사용하여 균주를 배양하는 단계를 포함하는 것으로 명시되는 경우, 수소는 배양에서 주요 에너지 공급원인 것으로 이해되어야 한다. 따라서, 질소 공급원으로 공급될 수 있는 암모니아 또는 소량의 유기 화합물과 같이 배양에서 다른 에너지 공급원이 소량 존재할 수 있지만, 상기 배양의 주요 대사작용 및 성장은 에너지 공급원으로서 수소의 이용에 기반한다. 전체 공정에서 수소는 바람직하게는 물 전기분해에 의해 생성되며; 즉, 전기로 물을 수소 및 산소 기체로 분해하여 생성된다. 따라서, 수소 및 산소 기체는 근처의 전기분해장치 (electrolyser)로부터 생물반응기로 제공된다. 대안으로서, 전극은 별도의 전기분해장치가 아닌 생물반응기에서 수소 및 산소를 생성하기 위해 생물반응기 내부에 배치될 수 있다.
이산화탄소를 포함하는 무기 탄소 공급원은 예를 들어 일산화탄소와 같은 다른 무기 탄소 공급원을 포함할 수 있다. 일 구체예에서, 기체 형태의 탄소 공급원 만이 배양에 제공된다. 바람직한 일 구체예에서, 이산화탄소는 배양에 제공되는 유일한 무기 탄소 공급원이며 실제로는 유일한 탄소 공급원이다. 일 구체예에서, 기체 및 미네랄 만이 배양에 제공되고, 기체 중 이산화탄소 수준은 10% 내지 50%, 예를 들어 15% 내지 45%, 예컨대 20% 내지 40%, 예를 들어, 25% 내지 35%, 예컨대 26% 내지 30%로 제공된다.
다른 구체예에서, 기체 및 미네랄이 배양에 제공되고, 기체 중 수소 (H2)의 수준은 30% 내지 80%, 예를 들어 35% 내지 75%, 예컨대 40% 내지 70%, 예를 들어 45% 내지 65%, 예컨대 50% 내지 60%로 제공된다.
다른 구체예에서, 기체 및 미네랄이 배양에 제공되고, 기체 중 산소 (O2)의 수준은 10% 내지 25%, 예를 들어 15% 내지 20%, 예컨대 16% 내지 18%로 제공된다. 다른 구체예에서, 산소 수준은 배양물 중 용존 산소 수준이 5% 내지 10%로 유지되도록 제공된다.
바람직한 일 구체예에서, 기체 및 미네랄 만이 배양으로 제공되며, H2, CO2 및 O2를 포함하는 기체가 제공되고, 여기서 H2의 퍼센트는 40% 내지 70%이며, CO2의 퍼센트는 18% 내지 28%이고, O2의 퍼센트는 12% 내지 22%이다.
전형적으로, 본 발명의 방법은 질소 공급원의 부가를 포함한다. 상기 질소 공급원은 예를 들어 수산화암모늄, 암모늄 염 예컨대 황산암모늄 또는 염화암모늄, 암모니아, 우레아 또는 니트레이트 예를 들어 질산칼륨의 형태로 제공될 수 있다. 다른 구체예에서, 질소 기체 (N2)가 질소 공급원으로서 제공된다. 바람직한 일 구체예에서, 상기 질소 공급원은 수산화암모늄 또는 암모늄 염 예컨대 황산암모늄이다.
일 구체예에서, 질소 공급원은 수산화 암모늄을 100 mg/L 내지 10 g/L, 예컨대 250 mg/L 내지 4 g/L, 예를 들어 0.5 g/L 내지 2 g/L, 예컨대 0.75 g/L 내지 1.5 g/L의 농도로 제공된다.
전형적으로, 본 발명의 방법은 미네랄, 예컨대 암모늄, 포스페이트, 칼륨, 나트륨, 바나듐, 철, 설페이트, 마그네슘, 칼슘, 몰리브덴, 망간, 붕소, 아연, 코발트, 셀레늄, 요오드, 구리 및/또는 니켈을 함유하는 미네랄의 부가를 포함한다. 적합한 미네랄 배지는 잘 알려진 기술이며, 예를 들어 Thermophilic Bacteria, CRC Press, Boca Raton, FL, Jacob K. Kristjansson, ed., 1992, for example on page 87, Table 4에 기재되었다.
일 구체예에서, 부가된 미네랄은 하기 중 하나 이상을 포함한다: 암모니아, 암모늄 (예: 염화암모늄 (NH4Cl), 황산암모늄 ((NH4)2SO4), 니트레이트 (예: 질산칼륨 (KNO3)), 우레아 또는 유기 질소 공급원; 포스페이트 (예: 인산이나트륨 (Na2HPO4), 인산칼륨 (KH2PO4), 인산 (H3PO4), 이티오인산칼륨 (K3PS2O2), 오르토인산칼륨 (K3PO4), 인산이나트륨 (Na2HPO4·2H2O) 인산이칼륨 (K2HPO4) 또는 인산일칼륨 (KH2PO4); 설페이트; 효모 추출물; 킬레이트화된 철 (예를 들어 EDTA 또는 시트르산으로 킬레이트화됨); 칼륨 (예: 인산칼륨 (KH2PO4), 질산칼륨 (KNO3), 요오드화칼륨 (KI), 브롬화칼륨 (KBr)); 및 기타 무기염, 미네랄 및 미량 영양소 (예: 염화나트륨 (NaCl), 황산마그네슘 (MgSO4·7H2O) 또는 염화마그네슘 (MgCl2), 염화칼슘 (CaCl2), 황산칼슘 (CaSO4) 또는 탄산칼슘 (CaCO3), 황산망간 (MnSO4·7H2O) 또는 염화망간 (MnCl2), 염화제2철 (FeCl2), 황산제1철 (FeSO4·7H2O) 또는 염화제1철 (FeCl2·4H2O), 중탄산나트륨 (NaHCO3) 또는 탄산나트륨 (Na2CO3), 황산아연 (ZnSO4) 또는 염화아연 (ZnCl2), 몰리브덴산암모늄 (NH4MoO4) 또는 몰리브덴산나트륨 (Na2MoO4·2H2O), 황산제1구리 (CuSO4) 또는 염화구리 (CuCl2·2H2O), 염화코발트 (CoCl2·6H2O) 또는 황산코발트 (CoSO4), 염화알루미늄 (AlCl3·6H2O), 염화리튬 (LiCl), 붕산 (H3BO3), 염화니켈 (NiCl2·6H2O) 또는 황산니켈 (NiSO4), 염화주석 (SnCl2·H2O), 염화바륨 (BaCl2·2H2O), 셀렌산구리 (CuSeO4·5H2O), 셀렌산나트륨 (Na2SeO4) 또는 아셀레늄산나트륨 (Na2SeO3), 메타바나데이트 나트륨 (NaVO3), 크롬 염).
바람직한 일 구체예에서, 본 발명의 방법은 하기 중 하나, 그 이상 또는 전부를 추가로 포함한다: NH4OH, KH2PO4, Na2HPO4·2H2O, NaVO3·H2O, FeSO4x7H2O, MgSO4·7H2O, CaSO4, Na2MoO4·2H2O, MnSO4·7H2O, ZnSO4·7H2O, H3BO3, CoSO4, CuSO4, NiSO4.
일 구체예에서, 세포에 제공된 배지는 1 g/L 미만의 클로라이드 염 (chloride salts), 예컨대 0.25 g/L 미만의 클로라이드 염, 예를 들어 0.1 g/L 미만의 클로라이드 염, 예컨대 0.025 g/L 미만의 클로라이드 염, 예를 들어 0.01 g/L 미만의 클로라이드를 포함한다. 일 구체예에서, 클로라이드 염은 배양에 공급되지 않는다.
다른 구체예에서, 공정 중에 비타민이 공급되지 않으며, 즉 배양에 제공된 배지는 비타민을 함유하지 않는다.
다른 구체예에서, 공정 중에 아미노산이 공급되지 않으며, 즉 배양에 제공된 배지는 아미노산을 함유하지 않는다.
다른 구체예에서, 공정 중에 유기 화합물이 공급되지 않으며, 즉 배양에 제공된 배지는 어떠한 유기 화합물도 함유하지 않는다.
소정의 구체예에서, 박테리아 배양물의 pH는 소정의 수준에서 제어된다. 소정의 구체예에서, pH는 박테리아 유지 및/또는 성장 및/또는 유기 화합물의 생산을 위한 최적 범위 내에서 제어된다. 일 구체예에서, 배양물의 pH는 5.5 내지 8.0, 예를 들어 6.5 내지 7.0, 예컨대 6.8로 유지된다.
소정의 구체예에서, 박테리아 배양물의 온도가 제어된다. 소정의 구체예에서, 온도는 박테리아 유지 및/또는 성장 및/또는 유기 화합물의 생산을 위한 최적 범위 내에서 제어된다. 일 구체예에서, 상기 배양물은 25℃ 내지 40℃, 예를 들어 28℃ 내지 32℃, 예컨대 30℃의 온도에서 성장한다.
전형적으로, 본 발명의 방법은 생물반응기에서 수행된다. 생물반응기가 세포 배양에 사용되며, 이들의 성장 곡선의 특정 단계에서 유지될 수 있다. 생물반응기의 사용은 화학독립영양 성장을 배양하는데 여러 면에서 유리하다. 일반적으로, 용존 이산화탄소, 산소, 및 다른 기체 예컨대 수소뿐만 아니라 다른 용존 영양소, 미량 원소, 온도 및 pH의 제어를 포함한 성장 조건의 제어가 생물반응기에서 용이하다. 기체 뿐만 아니라 영양 배지는 생물반응기에, 회분식 부가, 또는 주기적으로, 또는 검출된 고갈 또는 프로그래밍된 설정점에 대한 반응으로, 또는 배양물이 성장 및/또는 유지되는 기간 동안 연속적으로 부가될 수 있다. 연속 배양 공정에서, 기체 뿐만 아니라 영양 배지가 생물반응기에 연속적으로 부가된다. 또한, 박테리아-함유 배지는 생물반응기로부터 연속적으로 제거된다.
바람직한 일 구체예에서, 상기 박테리아 배양물의 부피는 100 mL 이상, 예컨대 1 L 이상, 예를 들어 10 L 이상, 예컨대 100 L 이상, 예를 들어 1,000 L 이상, 예컨대 10,000 L 이상, 예를 들어 50,000 L 이상, 예컨대 100,000 L 이상, 예를 들어 200,000 L 이상이다.
일 구체예에서, 배양물의 생산성은 시간당 리터당 0.1 g 초과의 세포 건조 중량, 예컨대 시간당 리터당 0.2 초과, 예를 들어 0.3 초과, 예컨대 0.4 초과, 예를 들어 0.5 초과, 예컨대 0.6 초과, 예를 들어 0.7 초과, 예컨대 0.8 초과, 예를 들어 0.9 초과, 예컨대 1 g 초과이다.
박테리아는 세포 은행 (cell bank)으로부터 직접, 또는 더 작은 규모로 시드 배양 (seed culture)을 통해 접종될 수 있다. 바람직하게는, 새로운 배지의 배양물로의 공급 및 소모된 배지와 박테리아의 제거는 생물반응기의 부피가 동일하게 유지되도록 동일한 속도로 발생한다.
일 구체예에서, 적절한 세포 밀도에 도달하는 초기 단계 후에, 박테리아는 이의 지수 단계 (log phase)로 연속적으로 유지되는 정상 상태 (steady state) 또는 유사 정상 상태 (pseudo steady state)로, OD600 5 이상, 예컨대 10 이상, 예를 들어 20 이상, 예컨대 50 내지 200, 예를 들어 50 내지 100으로 성장한다.
본 발명의 방법의 일 구체예에서, 상기 박테리아 균주는 0.04 - 0.12 h-1의 성장 속도를 갖는다.
본 발명의 방법의 다른 구체예에서, 연속상의 액체 공급 속도는 성장 속도의 50-80%이다.
크산토박터는 크산토박테라세에 (Xanthobacteraceae)과 유래의 그람-음성 박테리아의 속이다.
일 구체예에서, 본 발명의 방법에 사용된 크산토박터 균주는 이산화탄소를 생물체에 필수적인 유기 화합물, 예를 들어 글루코스로 전환하기 위해 Calvin Benson Bassham 경로를 사용하는 균주이다.
일 구체예에서, 본 발명의 방법에 사용된 크산토박터 균주는 수소 (H2)를 세포 에너지 등가물로 전환하기 위해 NiFeSe-하이드로게나제를 사용하는 균주이다.
일 구체예에서, 본 발명의 방법에 사용된 크산토박터 균주는 수소 (H2)를 세포 에너지 등가물로 전환하기 위해 NAD+-환원 하이드로게나제를 사용하는 균주이다.
일 구체예에서, 본 발명의 방법에 사용된 크산토박터 균주는 질소 고정이 가능하다.
일 구체예에서, 본 발명의 방법에 사용되는 박테리아 균주는 하기로 구성된 그룹으로부터 선택된다: X. 아길리스 (X. agilis), X. 아미녹시단스 (X. aminoxidans), X. 오토트로피쿠스 (X. autotrophicus), X. 플라부스 (X. flavus), X. 타게티디스 (X. tagetidis), X. 비스코서스 (X. viscosus), 크산토박터 종 126 (Xanthobacter sp. 126), 크산토박터 종 91 (Xanthobacter sp. 91) 및 균주 VTT-E-193585.
바람직한 일 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 VTT-E-193585 또는 X. 타게티디스 (X. tagetidis)이다. 가장 바람직하게는 본 발명의 방법에 사용되는 균주는 VTT-E-193585이다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 1에 제시된 16S 리보솜 RNA, 또는 서열번호: 1과 최대 20개, 예를 들어 1 내지 10개, 예컨대 1 내지 5개의 뉴클레오티드 차이, 예를 들어 서열번호: 1과 1개, 2개 또는 3개의 뉴클레오티드 차이를 갖는 16S 리보솜 RNA를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 3에 제시된 서열, 또는 서열번호: 3에 제시된 서열에 대해 93% 초과의 동일성, 예를 들어 95% 초과의 동일성, 예컨대 96% 초과의 동일성, 예를 들어 97% 초과의 동일성, 예컨대 98% 초과의 동일성, 예를 들어 99% 초과의 서열 동일성을 갖는 서열을 가진 리불로스-1,5-비스포스페이트 카복실라제/옥시게나제 (rubisco) 큰 사슬을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 5에 제시된 서열, 또는 서열번호: 5에 제시된 서열에 대해 83% 초과의 서열 동일성, 예를 들어 86% 초과의 동일성, 예컨대 90% 초과의 동일성, 예를 들어 95% 초과의 동일성, 예컨대 96% 초과의 동일성, 예를 들어 97% 초과의 동일성, 예컨대 98% 초과의 동일성, 예를 들어 99% 초과의 서열 동일성을 갖는 서열을 가진 리불로스-1,5-비스포스페이트 카복실라제/옥시게나제 (rubisco) 작은 사슬을 코딩하는 유전자를 포함한다.
서열번호: 2:
리불로스 비스포스페이트 카복실라제 큰 사슬의 뉴클레오티드 서열:
서열번호: 3:
리불로스 비스포스페이트 카복실라제 큰 사슬의 아미노산 서열
서열번호: 4:
리불로스 비스포스페이트 카복실라제 작은 사슬의 뉴클레오티드 서열:
서열번호: 5:
리불로스 비스포스페이트 카복실라제 작은 사슬의 아미노산 서열:
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 7에 제시된 서열, 또는 서열번호: 7에 제시된 서열에 대해 70% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 알파를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 9에 제시된 서열, 또는 서열번호: 9에 제시된 서열에 대해 77% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 베타를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 11에 제시된 서열, 또는 서열번호: 11에 제시된 서열에 대해 70% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 감마를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 13에 제시된 서열, 또는 서열번호: 13에 제시된 서열에 대해 79% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 델타를 코딩하는 유전자를 포함한다.
서열번호: 6:
NAD+-환원 하이드로게나제 HoxS 서브유닛 알파의 뉴클레오티드 서열:
서열번호: 7:
NAD+-환원 하이드로게나제 HoxS 서브유닛 알파의 아미노산 서열:
서열번호: 8:
NAD+-환원 하이드로게나제 HoxS 서브유닛 베타의 뉴클레오티드 서열:
서열번호: 9:
NAD+-환원 하이드로게나제 HoxS 서브유닛 베타의 아미노산 서열:
서열번호: 10:
NAD+-환원 하이드로게나제 HoxS 서브유닛 감마의 뉴클레오티드 서열:
서열번호: 11:
NAD+-환원 하이드로게나제 HoxS 서브유닛 감마의 아미노산 서열:
서열번호: 12:
NAD+-환원 하이드로게나제 HoxS 서브유닛 델타의 뉴클레오티드 서열:
서열번호: 13:
NAD+-환원 하이드로게나제 HoxS 서브유닛 델타의 아미노산 서열:
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 15에 제시된 서열, 또는 서열번호: 15에 제시된 서열에 대해 84% 초과의 서열 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NiFeSe 하이드로게나제 큰 서브유닛을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 17에 제시된 서열, 또는 서열번호: 17에 제시된 서열에 대해 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NiFeSe 하이드로게나제 작은 서브유닛을 코딩하는 유전자를 포함한다.
서열번호: 14:
주변세포질 [NiFeSe] 하이드로게나제 큰 서브유닛의 뉴클레오티드 서열:
서열번호: 15:
주변세포질 [NiFeSe] 하이드로게나제 큰 서브유닛의 아미노산 서열:
서열번호: 16:
주변세포질 [NiFeSe] 하이드로게나제 작은 서브유닛의 뉴클레오티드 서열:
서열번호: 17:
주변세포질 [NiFeSe] 하이드로게나제 작은 서브유닛의 아미노산 서열:
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 19에 제시된 서열, 또는 서열번호: 19에 제시된 서열에 대해 70% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 감마 사슬 atpG_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 21에 제시된 서열, 또는 서열번호: 21에 제시된 서열에 대해 78% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 알파 atpA_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 23에 제시된 서열, 또는 서열번호: 23에 제시된 서열에 대해 62% 초과의 동일성, 예를 들어 70% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 b atpF_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 25에 제시된 서열, 또는 서열번호: 25에 제시된 서열에 대해 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 c, 나트륨 이온 특이적 atpE_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 27에 제시된 서열, 또는 서열번호: 27에 제시된 서열에 대해 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 a atpB_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 29에 제시된 서열, 또는 서열번호: 29에 제시된 서열에 대해 71% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 엡실론 사슬 atpC_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 31에 제시된 서열, 또는 서열번호: 31에 제시된 서열에 대해 84% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 베타 atpD_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 33에 제시된 서열, 또는 서열번호: 33에 제시된 서열에 대해 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 베타 atpD_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 35에 제시된 서열, 또는 서열번호: 35에 제시된 서열에 대해 86% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 감마 사슬 atpG_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 37에 제시된 서열, 또는 서열번호: 37에 제시된 서열에 대해 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 알파 atpA_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 39에 제시된 서열, 또는 서열번호: 39에 제시된 서열에 대해 85% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 델타 atpH를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 41에 제시된 서열, 또는 서열번호: 41에 제시된 서열에 대해 87% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 b atpF_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 43에 제시된 서열, 또는 서열번호: 43에 제시된 서열에 대해 81% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 b' atpG_3을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 45에 제시된 서열, 또는 서열번호: 45에 제시된 서열에 대해 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 c atpE_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 47에 제시된 서열, 또는 서열번호: 47에 제시된 서열에 대해 92% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 a atpB_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 49에 제시된 서열, 또는 서열번호: 49에 제시된 서열에 대해 60% 초과의 동일성, 예를 들어 70% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 단백질 I atpI를 코딩하는 유전자를 포함한다.
서열번호: 18:
ATP 신타제 감마 사슬 atpG_1의 뉴클레오티드 서열
서열번호: 19:
ATP 신타제 감마 사슬 atpG_1의 아미노산 서열
서열번호: 20:
ATP 신타제 서브유닛 알파 atpA_1의 뉴클레오티드 서열
서열번호: 21:
ATP 신타제 서브유닛 알파 atpA_1의 아미노산 서열
서열번호: 22:
ATP 신타제 서브유닛 b atpF_1의 뉴클레오티드 서열
서열번호: 23:
ATP 신타제 서브유닛 b atpF_1의 아미노산 서열
서열번호: 24:
ATP 신타제 서브유닛 c, 나트륨 이온 특이적 atpE_1의 뉴클레오티드 서열
서열번호: 25:
ATP 신타제 서브유닛 c, 나트륨 이온 특이적 atpE_1의 아미노산 서열
서열번호: 26:
ATP 신타제 서브유닛 a atpB_1의 뉴클레오티드 서열
서열번호: 27:
ATP 신타제 서브유닛 a atpB_1의 아미노산 서열
서열번호: 28:
ATP 신타제 엡실론 사슬 atpC_1의 뉴클레오티드 서열
서열번호: 29:
ATP 신타제 엡실론 사슬 atpC_1의 아미노산 서열
서열번호: 30:
ATP 신타제 서브유닛 베타 atpD_1의 뉴클레오티드 서열
서열번호: 31:
ATP 신타제 서브유닛 베타 atpD_1의 아미노산 서열
서열번호: 32:
ATP 신타제 서브유닛 베타 atpD_2의 뉴클레오티드 서열
서열번호: 33:
ATP 신타제 서브유닛 베타 atpD_2의 아미노산 서열
서열번호: 34:
ATP 신타제 감마 사슬 atpG_2의 뉴클레오티드 서열
서열번호: 35:
ATP 신타제 감마 사슬 atpG_2의 아미노산 서열
서열번호: 36:
ATP 신타제 서브유닛 알파 atpA_2의 뉴클레오티드 서열
서열번호: 37:
ATP 신타제 서브유닛 알파 atpA_2의 아미노산 서열
서열번호: 38:
ATP 신타제 서브유닛 델타 atpH의 뉴클레오티드 서열
서열번호: 39:
ATP 신타제 서브유닛 델타 atpH의 아미노산 서열
서열번호: 40:
ATP 신타제 서브유닛 b atpF_2의 뉴클레오티드 서열
서열번호: 41:
ATP 신타제 서브유닛 b atpF_2의 아미노산 서열
서열번호: 42:
ATP 신타제 서브유닛 b' atpG_3의 뉴클레오티드 서열
서열번호: 43:
ATP 신타제 서브유닛 b' atpG_3의 아미노산 서열
서열번호: 44:
ATP 신타제 서브유닛 c atpE_2의 뉴클레오티드 서열
서열번호: 45:
ATP 신타제 서브유닛 c atpE_2의 아미노산 서열
서열번호: 46:
ATP 신타제 서브유닛 a atpB_2의 뉴클레오티드 서열
서열번호: 47:
ATP 신타제 서브유닛 a atpB_2의 아미노산 서열
서열번호: 48:
ATP 신타제 단백질 I atpI의 뉴클레오티드 서열
서열번호: 49:
ATP 신타제 단백질 I atpI의 아미노산 서열
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 51에 제시된 서열, 또는 서열번호: 51에 제시된 서열에 대해 60% 초과의 동일성, 예를 들어 70% 초과의 동일성, 예컨대 92% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 53에 제시된 서열, 또는 서열번호: 53에 제시된 서열에 대해 60% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 55에 제시된 서열, 또는 서열번호: 55에 제시된 서열에 대해 87% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_1을 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 57에 제시된 서열, 또는 서열번호: 57에 제시된 서열에 대해 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_2를 코딩하는 유전자를 포함한다.
다른 구체예에서, 본 발명의 방법에 사용된 박테리아 균주는 서열번호: 59에 제시된 서열, 또는 서열번호: 59에 제시된 서열에 대해 98.5% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 철 단백질 nifH를 코딩하는 유전자를 포함한다.
서열번호: 50:
니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_1의 뉴클레오티드 서열
서열번호: 51:
니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_1의 아미노산 서열
서열번호: 52:
니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_2의 뉴클레오티드 서열
서열번호: 53:
니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_2의 아미노산 서열
서열번호: 54:
니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_1의 뉴클레오티드 서열
서열번호: 55:
니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_1의 아미노산 서열
서열번호: 56:
니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_2의 뉴클레오티드 서열
서열번호: 57:
니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_2의 아미노산 서열
서열번호: 58:
니트로게나제 철 단백질 nifH의 뉴클레오티드 서열
서열번호: 59:
니트로게나제 철 단백질 nifH의 아미노산 서열
다운스트림 처리
일 구체예에서, 본 발명의 방법은 배양 중에 생산된 바이오매스를 수확하는 추가 단계를 포함한다. 바이오매스는 예를 들어 침강 (중력에 기반한 침전), 여과, 원심분리 또는 응집에 의해 수확될 수 있다. 응집에는 응집제의 부가가 필요할 수 있다. 원심분리는 예를 들어 연속 흐름 원심분리기를 사용하여 수행될 수 있다.
일 구체예에서, 수확된 바이오매스는 그 후에 건조된다. 건조는 예를 들어 원심분리, 드럼 건조, 증발, 동결 건조, 가열, 분무 건조, 진공 건조 및/또는 진공 여과를 포함하는 잘 알려진 방법을 사용하여 수행될 수 있다. 건조된 바이오매스는 그 후에 제품, 예를 들어 식품 또는 사료 제품 또는 사료 또는 식품 성분에 사용될 수 있다.
다른 구체예에서, 수확된 바이오매스의 세포가 용해된다. 일부 구체예에서 용해물은 불용성 분획 및 가용성 분획으로 분리될 수 있고, 이들 중 하나 또는 둘 모두는 그 후에 농축되거나 또는 건조될 수 있고, 후속하여 제품, 예를 들어 식품 또는 사료 제품에 사용될 수 있다.
일 구체예에서, 바이오매스가 수확되고, 상기 바이오매스로부터 단백질이 단리되어, 단백질 분획 및 비-단백질 성분을 포함하는 분획이 수득된다. 따라서, 일 구체예에서, 상기 방법은 단백질의 생산을 위한 것이고, 균주 VTT-E-193585 또는 이의 유도체를 배양하는 단계, 이어서 바이오매스를 수확하는 단계, 및 상기 바이오매스로부터 단백질을 단리하는 추가 단계를 포함한다. 다른 구체예에서, 상기 방법은 단백질의 생산을 위한 것이고, 에너지 공급원으로서 수소 및 무기 탄소 공급원을 사용하여 연속 배양으로 크산토박터 속의 박테리아 균주를 배양하는 단계 (여기서 무기 탄소 공급원은 이산화탄소를 포함함), 이어서 바이오매스를 수확하는 단계, 및 상기 바이오매스로부터 단백질을 단리하는 추가 단계를 포함한다. 단백질 단리 방법에 따라, 수득된 분획들은 더 순수하거나 또는 덜 순수할 수 있다. 따라서, 용어 "단백질 분획"은 단백질이 풍부한 분획을 의미한다. 단백질 분획은 여전히 상당한 양의 다른 성분을 포함할 수 있고, 또한 상당한 양의 단백질은 "비-단백질 성분을 포함하는 분획"이 될 수 있다.
단백질의 단리는 임의의 적절한 방법을 사용하여 수행될 수 있다. 예를 들어, 일 구체예에서, 단백질은 세포를 기계적으로 파괴하고, 단백질을 세포 데브리스로부터 하나 이상의 여과 단계, 예를 들어 감소하는 공극 크기를 가진 다중 필터를 통한 연속 여과를 통해 분리함으로써 단리된다. 기계적 파괴는 임의의 적절한 방법 예를 들어 볼 밀링, 초음파 처리, 균질화, 고압 균질화, 기계적 전단 등을 사용하여 수행될 수 있다. 수득된 여과된 단백질 분획은 단백질이 풍부하지만, 여전히 다른 더 작은 성분들을 함유할 것이다. 단백질은 선택적으로 이러한 분획으로부터 임의의 적절한 방법을 사용하여 추가로 정제될 수 있다.
다른 구체예에서, 단백질 분획은 에탄올 추출을 수행한 다음에 1회 이상의 여과 단계를 수행함으로써 단리된다. 이러한 방법은 예를 들어 대두 단백질의 제조로부터 알려져 있다 (예를 들어, Chapter 5 "Soybean Protein Concentrates" in "Technology of production of edible flours and protein products from soybeans" by Berk FAO Agricultural Services Bulletin No. 97 (1992) 참조). 수득된 단백질 분획은 단백질이 풍부하지만, 여전히 다른 성분을 함유할 것이다. 단백질은 선택적으로 이러한 분획으로부터 임의의 적절한 방법을 사용하여 추가로 정제될 수 있다.
일 구체예에서, 본 발명의 방법은 본 발명의 방법으로부터 수득된 단백질 분획을 가수분해하여 아미노산 및 작은 펩티드를 수득하는 추가 단계를 포함한다.
본 발명의 방법의 일 구체예에서, 상기 방법은 상기 바이오매스, 상기 단백질 분획, 또는 상기 비-단백질 성분을 포함하는 분획으로부터 식품 또는 사료 제품을 제조하는 추가 단계를 포함한다. 상기 추가 단계는 단순히 상기 바이오매스, 단백질 분획 또는 비-단백질 성분을 포함하는 분획을 식품 또는 사료 제품의 제조 중에 부가함으로써, 식품 또는 사료 제품에 혼입시키는 단계를 포함할 수 있다. 다른 구체예에서, 바이오매스 또는 이의 분획의 추가 정제 또는 변형이 이의 식품 또는 사료 제품으로의 혼입 과정 중에 수행된다.
추가 양상에서, 본 발명은 본 발명에 따른 방법에 의해 수득되거나 또는 수득 가능한 산물, 예컨대 바이오매스, 단백질 또는 비-단백질 성분에 관한 것이다.
일 구체예에서, 본 발명의 방법으로부터 수득된 산물은 40% 초과의 단백질, 예를 들어 40% 내지 99%의 단백질, 예를 들어 40% 내지 90%의 단백질, 예를 들어 40% 내지 60%의 단백질을 포함한다. 특정 구체예에서, 산물은 25% 내지 75%의 단백질, 0% 내지 20%의 지질 및 5% 내지 40%의 탄수화물을 포함한다. 추가 구체예에서, 산물은 40% 내지 60%의 단백질, 0% 내지 15%의 지질 및 10% 내지 25%의 탄수화물을 포함한다. 또 다른 구체예에서, 본 발명의 방법으로부터 수득된 산물은 45% 내지 55%의 단백질, 5% 내지 10%의 지질 및 10% 내지 20%의 탄수화물을 포함한다.
상기 기재된 바와 같이, 추가 양상에서 본 발명은 본 발명에 따른 방법에 의해 수득되거나 또는 수득 가능한 식품 또는 사료 제품에 관한 것이다. 용어 "식품" 및 "사료"가 본원에서 사용되는 경우, 가공 식품과 같은 기존의 식품 및 사료 제품뿐만 아니라, 단백질 바, 분말 또는 쉐이크와 같은 식품 및 사료 보충제, 육류 대체품, 식품 성분, 프로바이오틱스, 프리바이오틱스, 기능식품 등과 같은 관련 제품을 포함하는 것으로 의도된다. 소정의 구체예에서, 상기 바이오매스, 상기 단백질 분획 또는 상기 비-단백질 성분을 포함하는 분획은 베지테리언 (vegetarian) 또는 비건 (vegan) 식품의 생산에 이용된다.
본 발명은 하기의 비-제한적인 실시예를 사용하여 추가로 예시된다:
실시예
실시예 1. 화학독립영양 성장이 가능한 박테리아 균주의 단리
핀란드 Naantali의 발트해 해변에서 토양과 해수를 함유하는 50 mL의 샘플을 멸균 falcon 튜브에 수집하였다. 토양 샘플의 일부를 멸균 삼각 플라스크내에서 10 mL의 미네랄 배지와 혼합하였다. 상기 배지는 수돗물 중에서 제조된 1 g/L NH4OH, 0.23 g/L KH2PO4, 0.29 g/L Na2HPO4·2 H2O, 0.005 g/L NaVO3·H2O, 0.2 g/L FeSO4·7 H2O, 0.5 g/L MgSO4·7 H2O, 0.01 g/L CaSO4, 0.00015 g/L Na2MoO4·2 H2O, 0.005 g/L MnSO4, 0.0005 g/L ZnSO4·7 H2O, 0.0015 g/L H3BO3, 0.001 g/L CoSO4, 0.00005 g/L CuSO4 및 0.0001 g/L NiSO4로 구성되었다. 토양 및 배지의 현탁액을, 150 mL/min의 N2, 18 mL/min의 H2, 3 mL/min의 O2 및 6 mL/min의 CO2의 기체 혼합물로 연속적으로 플러싱된 밀봉된 강철 박스내에서 +30℃ 온도의 진탕 인큐베이터에서 인큐베이션하였다. 7일 간격으로 1 mL의 현탁액을 채취하여 상기 배양을 재충전하고, 이를 멸균 조건에서 삼각 플라스크내의 배지 9 mL에 부가한 다음에, 다시 인큐베이션 박스에 배치하였다. 네 번째 희석 후에, 현탁액 중에 눈에 띄는 토양이 남아 있지 않았다. 생물반응기 배양을 위해 바이오매스를 증가시키기 위해 세포 현탁액의 부피를 100 mL로 증가시켰다. 상기 현탁액이 15-용기 200-mL 병렬 생물반응기 시스템 (Medicel Explorer, Medicel Oy, Finland)에서 190 mL의 미네랄 배지에 접종되었을 때 상기 현탁액의 광학 밀도 (OD600)는 1.53이었다. 배양 조건은 800 rpm 교반, +30℃ 온도 및 pH는 1M NaOH로 제어하여 6.8로 설정하였다. 14 mL/min H2, 3 mL/min O2 및 6 mL/min CO2로 구성된 기체 혼합물을 갖는 스퍼저 (sparger)를 통해 기체를 공급하였다. 반응기의 헤드 공간을 300 mL/min 공기로 플러싱하였다. 연속 배양에 미네랄 배지를 6 mL/h로 공급하였고, 세포 현탁액을 반응기로부터 모세관을 통해 빼내어 부피를 200 mL로 일정하게 유지하였다. 반응기로부터 채취한 세포 현탁액을 +4℃에서 저장하였다. 생물반응기로부터 샘플을 매일 자동으로 채취하고, 600 nm에서 흡광도를 측정하여 성장을 모니터링하였다. 생물반응기에서 498시간 배양 후에, 샘플을 무균 상태로 채취하고, 현탁액을 희석하여, 상기 미네랄 및 2% 박테리아용 아가 (bacteriological agar)를 함유하는 아가 미네랄 배지 플레이트에 플레이팅하였다. 삼각 플라스크의 경우에 대해 상기 기재된 바와 동일한 조건에서 플레이트를 인큐베이션하였다. 그 다음에 아가 플레이트로부터 콜로니를 선택하고, 하나의 콜로니에서 하나의 유기체를 단리하기 위해 새로운 아가 플레이트에 스트리킹하였다. 이를 2회 반복하였다. 단일 콜로니를 선택하고, 96-웰 마이크로타이터 플레이트에 있는 200 μL의 배지에 현탁시켰다. 현탁액을 +30℃ 온도, 및 150 mL/min의 N2, 18 mL/min의 H2, 3 mL/min의 O2 및 6 mL/min의 CO2로 연속적으로 플러싱된 EnzyScreen 기밀 박스에서 625 rpm으로 진탕하면서 인큐베이션하였다. 하나의 웰의 현탁액을 삼각 플라스크로 옮기고, 새로운 배지로 보충하였다. 생물반응기 배양을 수행하기에 충분한 바이오매스가 생산될 때까지 부피를 증가시켰다. 유기체는 VTT-E-193585로 VTT culture collection에 기탁하였다.
샘플의 16S rRNA 시퀀싱으로 상기 샘플은 단 하나의 유기체를 함유하였음을 보여주었다. 1x150 bp 메타게놈 샷건 서열 (metagenomic shotgun sequences)을 제공하는 Illumina NextSeq 시퀀싱에 동일한 샘플을 사용하였다. Unicycler (Wick et al, 2017 PLoS computational biology 13:e1005595)를 사용하여, 101개의 contig로 구성된 메타게놈 서열에 대한 데 노보 (de novo) 조립체를 만들었다. 전체 게놈 길이는 4,846,739 bp이었고, GC 함량은 67.9%이었다. 유전자 예측 및 기능 주석은 Prokka를 사용하여 수행하였다 (Seemann, 2014 Bioinformatics 30:2068). 게놈 주석은 4,429개의 유전자를 생성하였다. Roary pan 게놈 정렬 (Page et al, 2015 Bioinformatics 31:3691)은 크산토박터 종 중에서 VTT-E-193585를 그룹화하였다. 그러므로 상기 균주는 크산토박터 종으로 동정되었고, 가장 가까운 게놈은 크산토박터 타게티디스 (Xanthobacter tagetidis)이다. 이종상동성 단편 (orthologous fragments)만을 고려한 평균 뉴클레오티드 동일성의 정렬-기반 계산 (OrthoANI) (Lee et al, 2016 Int J Syst Evol Microbiol 66:1100)으로 크산토박터 타게티디스 (ATCC 700314; GCF_003667445.1)에 대해 80.4%로 최고 일치를 제공하였고, 제시된 종 경계 컷-오프 (species boundary cut-off)는 95-96%이다 (예를 들어, Chun et al., 2018 Int J Syst Evol Microbiol, 68: 461-466 참조). 크산토박터 오토트로피쿠스 Py2는 79.6%의 일치율을 제공한 반면에, 크산토박터 종 91에 대한 일치율은 79.0%이었다. 따라서 VTT-E-193585로 기탁된 단리된 박테리아 균주는 프로테오박테리아 (Proteobacteria) 문; 알파 프로테오박테리아 (Alpha Proteobacteria) 강; 리조비알레스 (Rhizobiales) 속에 속한다고 결론내릴 수 있었다. 가장 가능성이 높은 과 (Family)는 크산토박테라세에 (Xanthobacteraceae), 속은 크산토박터 (Xanthobacter)이다. VTT-E-193585 박테리아 균주는 임의의 알려진 종에 명확하게 배정할 수 없었다.
추정되는 항미생물제 내성 유전자에 대한 검색을 수행하였다. ABRicate (https://github.com/tseemann/abricate) 툴은 blastn 또는 blastp를 사용하여 Arg-Annot, NCBI, ResFinder, the ecOH, Megares 및 VFDB 데이터베이스에 대해 게놈을 검색하는데 사용하였다. 뉴클레오티드 및 단백질 수준 모두에서 동일성 및 적용 범위 모두에 대해 50%의 역치 (threshold)를 설정하였다. 추정되는 항미생물제 내성 유전자 2개만이 확인되었다. 이러한 2개의 유전자는 항생제 내성과 관련된 아미노산 변화를 함유하지 않았으므로, 내성 표현형은 예상되지 않는다.
실시예 2. 단리된 박테리아 균주의 파일럿 배양 및 분석
VTT-E-193585로 기탁된 단리된 박테리아 균주를 기존의 200-리터 교반 탱크 생물반응기 (MPF-U, Marubishi Ltd, Japan)에서 배양하였다. 400 rpm으로 회전하는 Rushton-타입 임펠러를 사용하여 혼합을 수행하였다. 배양 온도는 +30℃로 유지하였다. 소프트웨어 제어에 의해 8M NaOH 또는 3.6M H3PO4를 부가하여 pH를 6.8±0.2로 유지하였다. 배양 배지는 수돗물 중에서 제조된 1 g/L NH4OH, 0.23 g/L KH2PO4, 0.29 g/L Na2HPO4·2 H2O, 0.005 g/L NaVO3·H2O, 0.2 g/L FeSO4·7 H2O, 0.5 g/L MgSO4·7 H2O, 0.01 g/L CaSO4, 0.00015 g/L Na2MoO4·2 H2O, 0.005 g/L MnSO4, 0.0005 g/L ZnSO4·7 H2O, 0.0015 g/L H3BO3, 0.001 g/L CoSO4, 0.00005 g/L CuSO4 및 0.0001 g/L NiSO4를 함유하였다. 1.8-10.5 L/min의 수소 기체, 0.6-2.5 L/min의 산소 기체 및 1.8-5 L/min의 이산화탄소 기체를 함유하는 혼합물을 주요 에너지 및 탄소 공급원으로 일정하게 공급하였다. 기체 혼합물 조성을 조정하여 용존 산소 수준을 7.2±0.5%로 유지하였다. 배양을 위한 접종원은 실시예 1에 기재된 바와 같이 제조하였다. 샘플을 수동으로 채취하고, 600 nm에서 흡광도 (Ultrospec 2100 pro UV/visible spectrophotometer, Biochrom Ltd., England)를 측정하여 광학 밀도로서 세포 밀도를 분석하고, 105℃의 오븐에서 밤새 건조시켜서 세포 건조 중량 (CDW)을 측정하여 성장을 모니터링하였다. 광학 밀도는 또한 인 시추 (in situ) 흡광도 프로브 (Trucell 2, Finesse Ltd, USA)를 사용하여 모니터링하였다. 배양의 성장 곡선을 도 1에 제시하였다. 회분식 단계에서 최대 성장 속도는 0.06 h-1이었다. 최대 세포 밀도는 92시간에 4.5 g_CDW/L이었다. 92시간의 배양 후에, 상기 기재된 신선한 배양 배지의 공급을 0.01 h-1의 희석율로 개시하였다. 공급을 계속하는 동안, 세포 밀도는 평균 2.9 g_CDW/L이었다. 배양액을 냉각된 (+10℃) 탱크에 일정하게 수집하였고, 이 탱크로부터 연속 원심분리기 (BTPX-205, Alfa-Laval AB, Sweden)에 300-리터 배치로 공급하였다. 상기 분리기로부터 수집된 세포-함유 슬러리를 대기압 이중 드럼 건조기 (Buflovak 6x8 ADDD, Hebeler process solutions Llc., USA)로 공급하였고, 이를 4 bar의 스팀으로 가열하였고, 드럼은 3.5 rpm으로 회전하였다. 그 결과 건조 물질 함량이 약 96%인 건조 세포 분말을 수득하였다. 건조 세포 분말의 분석 결과를, 표 1에 근사 조성 (proximate composition), 표 2에 아미노산 조성, 표 3에 지방산 조성, 표 4에 비타민 함량을 제시하였다. 분석에 따르면 건조 세포 분말은 모든 필수 아미노산과 함께 높은 단백질 함량을 갖는 것으로 나타났다. 이는 또한 포화 지방산보다 불포화 지방산을 더 많이 함유하였고, 비타민 B군을 많이 함유하였다. 펩티도글리칸 함량은 단지 0.002 mg/g_CDW이었고, 리포폴리사카라이드 함량은 0.01 mg/g_CDW이었다. 이들 농도는 가능한한 적은 것이 유리할 것이다. 비교로서, 동시에 상업용 락트산 박테리아 제제를 분석하여, 펩티도글리칸 함량은 0.244 mg/g_DW이었고, 리포폴리사카라이드 함량은 0.015 mg/g_DW이었다. 세포독성 및 유전독성 분석은 배양 상등액 샘플을 사용하여 수행하였다. HepG2 또는 HeLa229 인간 세포주에 대한 세포독성은 관찰되지 않았다. 대장균 (Escherichia coli) WP2 trp- 또는 CM871 uvrA recA lexA 균주에 대한 유전독성은 관찰되지 않았다.
파라미터 | 방법 | 단위 | 값 |
수분 | 103 ℃에서 건조 | g/100g | 3.3 |
단백질 | Kjeldahl (N x 6,25) | g/100g | 72.2 |
지방 | Weibull-Stoldt | g/100g | 6.0 |
포화 지방산 | 표 3에 기반하여 산출 | g/100g | 1.8 |
일-불포화 지방산 | 표 3에 기반하여 산출 | g/100g | 3.8 |
다중불포화 지방산 | 표 3에 기반하여 산출 | g/100g | 0.4 |
오메가 3 지방산 | 표 3에 기반하여 산출 | g/100g | < 0.01 |
오메가 6 지방산 | 표 3에 기반하여 산출 | g/100g | 0.4 |
식이섬유 | 중량 측정 | g/100g | 10.5 |
회분 | 550 ℃에서 회분화 | g/100g | 5.8 |
글루코스 | HPLC-ELSD | g/100g | < 0.15 |
프럭토스 | HPLC-ELSD | g/100g | < 0.1 |
수크로스 | HPLC-ELSD | g/100g | < 0.1 |
락토스 | HPLC-ELSD | g/100g | < 0.25 |
말토스 | HPLC-ELSD | g/100g | < 0.2 |
전체 당 | 산출 | g/100g | < 0.8 |
탄수화물 | 산출 | g/100g | 2.2 |
에너지 | 산출 | kJ/100g | 1572 |
에너지 | 산출 | kcal/100g | 373 |
파라미터 | 방법 | 단위 | 값 |
리신 | 이온 크로마토그래피 | % | 3.95 |
메티오닌 | 이온 크로마토그래피 | % | 1.60 |
시스테인 | 이온 크로마토그래피 | % | 0.39 |
아스파르트산 | 이온 크로마토그래피 | % | 6.82 |
트레오닌 | 이온 크로마토그래피 | % | 3.47 |
세린 | 이온 크로마토그래피 | % | 2.75 |
글루탐산 | 이온 크로마토그래피 | % | 8.84 |
프롤린 | 이온 크로마토그래피 | % | 3.14 |
글리신 | 이온 크로마토그래피 | % | 4.40 |
알라닌 | 이온 크로마토그래피 | % | 6.94 |
발린 | 이온 크로마토그래피 | % | 4.96 |
이소류신 | 이온 크로마토그래피 | % | 3.34 |
류신 | 이온 크로마토그래피 | % | 6.08 |
티로신 | 이온 크로마토그래피 | % | 2.99 |
페닐알라닌 | 이온 크로마토그래피 | % | 4.58 |
히스티딘 | 이온 크로마토그래피 | % | 1.66 |
아르기닌 | 이온 크로마토그래피 | % | 4.96 |
트립토판 | HPLC | % | 1.34 |
파라미터 | 방법 | 단위 | 값 |
C16:0 (팔미트산) | GC-MS | % | 24.8 |
C16:1 (팔미톨레산) | GC-MS | % | 3.0 |
C18:0 (스테아르산) | GC-MS | % | 4.4 |
C18:1n9 (올레산) | GC-MS | % | 59.9 |
C18:2n6 (리놀레산) | GC-MS | % | 6.1 |
C18:3n3 (알파-리놀렌산) | GC-MS | % | 0.4 |
파라미터 | 방법 | 단위 | 값 |
비타민 A (RE) | HPLC | IU/100 g | <100 |
비타민 E (TE) | HPLC | mg/100 g | 0.33 |
비타민 D3 | HPLC | IU/100 g | <10 |
비타민 D2 | HPLC | IU/100 g | 21.6 |
비타민 C | HPLC | mg/100 g | <1 |
티아민 클로라이드 염산염 | LC-MS/MS | mg/100 g | 0.9 |
비타민 B1 (티아민) | 산출 | mg/100 g | 0.708 |
비타민 B2 (리보플라빈) | HPLC | mg/100 g | 6.27 |
피리독신 염산염 | HPLC | mg/100 g | 3.39 |
비타민 B6 (피리독신) | 산출 | mg/100 g | 2.79 |
비타민 B12 | LC-MS/MS | μg/100 g | 224 |
콜린 클로라이드 | LC-MS/MS | mg/100 g | 14.3 |
바이오틴 | LC-MS/MS | μg/100 g | 15.6 |
엽산 | Microbiological | μg/100 g | 1270 |
니아신 (비타민 B3) | Microbiological | mg/100 g | 23.2 |
판토텐산 | Microbiological | mg/100 g | 6.53 |
실시예 3. 다양한 질소 공급원에서 단리된 박테리아 균주의 배양.
VTT-E-193585로 기탁된 단리된 박테리아 균주를 15-용기 병렬 생물반응기 시스템에서 200 mL 부피 (Medicel Explorer, Medicel Oy, Finland)로 배양하였다. 800 rpm으로 회전하는 Rushton-타입 임펠러를 사용하여 혼합을 수행하였다. 배양 온도는 +30℃로 유지하였다. 1M NaOH를 부가하여 pH를 6.8로 유지하였다. 배양 배지는 수돗물 중에서 제조된 0.23 g/L KH2PO4, 0.29 g/L Na2HPO4·2 H2O, 0.005 g/L NaVO3·H2O, 0.2 g/L FeSO4·7 H2O, 0.5 g/L MgSO4·7 H2O, 0.01 g/L CaSO4, 0.00015 g/L Na2MoO4·2 H2O, 0.005 g/L MnSO4, 0.0005 g/L ZnSO4·7 H2O, 0.0015 g/L H3BO3, 0.001 g/L CoSO4, 0.00005 g/L CuSO4 및 0.0001 g/L NiSO4를 함유하였다. 또한, 4개의 배양은 18.7 mM NH4OH를 함유하고, 4개의 배양은 9.34 mM 우레아 (OC(NH2)2)를 함유하며, 4개의 배양은 18.7 mM의 질산칼륨 (KNO3)을 함유하고, 3개의 배양은 배지 중에 질소 공급원 없이 유지되도록, 배양에서 질소 공급원을 가변하였다. 22 mL/min의 수소 기체, 3.2 mL/min의 공기 및 6.4 mL/min의 이산화탄소를 함유하는 혼합물을 주요 에너지 및 탄소 공급원으로 일정하게 공급하였다. 따라서, 공기와 함께, 모든 배양에는 또한 질소 기체를 공급하였다. 샘플을 자동으로 채취하고, 600 nm에서 흡광도 (Ultrospec 2100 pro UV/visible spectrophotometer, Biochrom Ltd., England)를 측정하여, 광학 밀도로서 세포 밀도를 분석하여 성장을 모니터링하였다. 배양의 성장 곡선을 도 2에 제시하였다. 암모니아 및 우레아에서의 성장은 유사하였다. 니트레이트 또는 질소 기체에서의 성장은 암모니아 또는 우레아에서보다 명확하게 더 느렸다. 배양 종료 무렵, 니트레이트에서의 성장은 유일한 질소 공급원으로서 질소 기체에서의 성장보다 더 우수하였다. 그럼에도 불구하고 질소 기체가 유일한 질소 공급원인 배양에서도 성장하였고, 이는 VTT-E-193585로 기탁된 단리된 박테리아 균주가 질소 고정이 가능함을 보여주었다.
실시예 4. 항생제 감수성의 특성규명
VTT-E-193585로 기탁된 단리된 박테리아 균주에 대한 겐타마이신, 카나마이신, 스트렙토마이신, 테트라사이클린, 암피실린, 시프로플록사신, 콜리스틴 및 포스포마이신의 항생제 감수성은, 양이온-조정된 Mueller Hinton Broth -배지 (LabM, LAB114, 양이온 Mg2+ 및 Ca2+ 각각 부가)를 사용하여 +35 ± 2℃의 호기성 조건에서 48 ± 1시간 동안, 포스포마이신의 경우 아가 희석 방법 (agar dilution method)을 사용하고, 겐타마이신, 카나마이신, 스트렙토마이신 및 테트라사이클린의 경우 브로스 미세희석 방법 (broth microdilution method)을 사용하는, VetMIC Lact-1 플레이트 (SVA National Veterinary Institute, Uppsala, Sweden)에서, 암피실린, 시프로플록사신 및 콜리스틴의 경우 핸드-메이트 미세희석 플레이트 (hand-made microdilution plate)에서, CLSI M07-A111 표준 (Clinical and laboratory standards institute. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically, 11th ed. CLSI standard M07, 2018)에 따라 분석하였다. 대장균 ATCC 25922는 품질 대조 균주로서 사용하였고, 이는 호기성 조건에서 +35 ± 2℃에서 18 ± 2시간 동안 인큐베이션하였다. 균주들의 항생제 감수성 결과를 표 5에 제시하였다. 단리 박테리아 균주는 일반적으로 항생제에 민감한 것으로 밝혀졌다. 겐타마이신, 카나마이신, 스트렙토마이신 및 테트라사이클린의 경우, VTT-E-193585에 대한 최소 억제 농도 (MIC) 값은 대장균 ATCC 25922에 비해 낮거나 또는 비슷하였지만, 암피실린, 시프로플록사신, 콜리스틴 및 포스포마이신의 경우, MIC 값은 VTT-E-193585에서 더 높았다.
VTT-E-193585
48 h ± 1 h |
대장균 ATCC 25922
18 h ± 2 h |
|
겐타마이신 | 0.5 | 0.5 |
카나마이신 | 2 | 4 |
스트렙토마이신 | 0.5 | 4 |
테트라사이클린 | 0.25 | 1 |
암피실린 | 16 | 8 |
시프로플록사신 | 0.06 | 0.008 |
콜리스틴 | 4 | 2 |
포스포마이신 | 32 | 0.5 |
SEQUENCE LISTING
<110> Solar Foods
<120> STRAINS AND PROCESSES FOR SINGLE CELL PROTEIN OR BIOMASS
PRODUCTION
<130> Solar002
<160> 59
<170> PatentIn version 3.5
<210> 1
<211> 1482
<212> DNA
<213> Unknown
<220>
<223> Novel strain
<400> 1
cttgagagtt tgatcctggc tcagagcgaa cgctggcggc aggcctaaca catgcaagtc 60
gagcgcccag caatgggagc ggcagacggg tgagtaacgc gtggggatgt gcccaatggt 120
acggaataac ccagggaaac ttggactaat accgtatgag cccttcgggg gaaagattta 180
tcgccattgg atcaacccgc gtctgattag ctagttggtg gggtaacggc ccaccaaggc 240
gacgatcagt agctggtctg agaggatgat cagccacact gggactgaga cacggcccag 300
actcctacgg gaggcagcag tggggaatat tggacaatgg gcgcaagcct gatccagcca 360
tgccgcgtgt gtgatgaagg ccttagggtt gtaaagcact ttcgccggtg aagataatga 420
cggtaaccgg agaagaagcc ccggctaact tcgtgccagc agccgcggta atacgaaggg 480
ggctagcgtt gctcggaatc actgggcgta aagcgcacgt aggcggatcg ttaagtcagg 540
ggtgaaatcc tggagctcaa ctccagaact gcccttgata ctggcgacct tgagttcgag 600
agaggttggt ggaactgcga gtgtagaggt gaaattcgta gatattcgca agaacaccag 660
tggcgaaggc ggccaactgg ctcgatactg acgctgaggt gcgaaagcgt ggggagcaaa 720
caggattaga taccctggta gtccacgccg taaacgatgg atgctagccg ttgggcagct 780
tgctgttcag tggcgcagct aacgcattaa gcatcccgcc tggggagtac ggtcgcaaga 840
ttaaaactca aaggaattga cgggggcccg cacaagcggt ggagcatgtg gtttaattcg 900
aagcaacgcg cagaacctta ccagcctttg acatggcagg acgatttcca gagatggatc 960
tcttccagca atggacctgc acacaggtgc tgcatggctg tcgtcagctc gtgtcgtgag 1020
atgttgggtt aagtcccgca acgagcgcaa ccctcgcctc tagttgccag cattcagttg 1080
ggcactctag agggactgcc ggtgataagc cgagaggaag gtggggatga cgtcaagtcc 1140
tcatggccct tacgggctgg gctacacacg tgctacaatg gtggtgacag tgggatgcga 1200
aagggcgacc tctagcaaat ctccaaaagc catctcagtt cggattgtac tctgcaactc 1260
gagtgcatga agttggaatc gctagtaatc gtggatcagc atgccacggt gaatacgttc 1320
ccgggccttg tacacaccgc ccgtcacacc atgggagttg gctttacccg aaggcgctgc 1380
gctaacccgc aagggaggca ggcgaccacg gtagggtcag cgactggggt gaagtcgtaa 1440
caaggtagcc gtaggggaac ctgcggctgg atcacctcct tt 1482
<210> 2
<211> 1467
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 2
atgggtgccg aagcaaccgt cgggcagatc acggacgcca agaagagata cgccgccggc 60
gtgctgaagt acgcccagat gggctactgg aacggcgact acgttcccaa ggacaccgac 120
ctcctggcgg tgttccgcat caccccccag gcgggcgtgg acccggtgga agccgccgcg 180
gcggtcgccg gcgaaagctc caccgctacc tggaccgtgg tgtggaccga ccggctcacc 240
gccgccgacg tctaccgcgc caaggcctac aaggtggagc cggtgccggg ccaggaaggc 300
cagtatttct gctacatcgc ctatgatctc gatttgttcg aggaaggctc catcgccaac 360
ctcacggcgt cgatcatcgg caacgtcttc tccttcaagc cgctgaaggc ggcgcggctg 420
gaggacatgc ggcttcccgt cgcctatgtg aagaccttcc gcggcccgcc caccggcatc 480
gtggtcgagc gcgagcgcct ggacaagttc ggccgccccc ttctgggcgc caccaccaag 540
ccgaagcttg gcctctcggg caagaattac ggccgcgtgg tctatgaggc cctcaagggc 600
ggcctcgact tcgtgaagga cgacgagaac atcaactcgc agcccttcat gcactggcgc 660
gatcgcttcc tctattgcat ggaggccgtc aacaaggccc aggccgagac cggcgaggtg 720
aaggggcact atctcaacat caccgccggg accatggagg agatgtaccg ccgcgccgag 780
ttcgccaagg aactgggctc cgtggtggtg atggtggatc tcatcatcgg ctggaccgcc 840
atccagtcca tgtccaactg gtgccgcgag aacgacatga tcctgcacat gcaccgtgcg 900
ggccatggca cctacacgcg ccagaagagc cacggcgtct ccttccgcgt catcgccaag 960
tggctgcggc tcgccggcgt cgaccacctg cacaccggca ccgccgtggg caagctggaa 1020
ggcgacccca tgaccgtgca gggcttctac aatgtctgcc gcgagacgac gacgcagcag 1080
gacctcaccc gcggcctgtt cttcgagcag gactggggcg gcatccgcaa ggtgatgccg 1140
gtggcctccg gcggcatcca tgcgggccag atgcaccagc tcatcgacct gttcggcgag 1200
gacgtggtgc tccagttcgg cggcggcacc atcggccacc cggacggcat ccaggccggc 1260
gccaccgcca accgcgtggc gctggaaacc atgatcctcg cccgcaacga gggccgcgac 1320
atcaggaacg agggcccgga aatcctggtg gaagccgcca aatggtgccg tccgctgcgc 1380
gcggcgctcg atacctgggg cgaggtgacc ttcaactacg cctccaccga cacgtccgat 1440
tacgtgccca ccgcgtccgt cgcctga 1467
<210> 3
<211> 488
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 3
Met Gly Ala Glu Ala Thr Val Gly Gln Ile Thr Asp Ala Lys Lys Arg
1 5 10 15
Tyr Ala Ala Gly Val Leu Lys Tyr Ala Gln Met Gly Tyr Trp Asn Gly
20 25 30
Asp Tyr Val Pro Lys Asp Thr Asp Leu Leu Ala Val Phe Arg Ile Thr
35 40 45
Pro Gln Ala Gly Val Asp Pro Val Glu Ala Ala Ala Ala Val Ala Gly
50 55 60
Glu Ser Ser Thr Ala Thr Trp Thr Val Val Trp Thr Asp Arg Leu Thr
65 70 75 80
Ala Ala Asp Val Tyr Arg Ala Lys Ala Tyr Lys Val Glu Pro Val Pro
85 90 95
Gly Gln Glu Gly Gln Tyr Phe Cys Tyr Ile Ala Tyr Asp Leu Asp Leu
100 105 110
Phe Glu Glu Gly Ser Ile Ala Asn Leu Thr Ala Ser Ile Ile Gly Asn
115 120 125
Val Phe Ser Phe Lys Pro Leu Lys Ala Ala Arg Leu Glu Asp Met Arg
130 135 140
Leu Pro Val Ala Tyr Val Lys Thr Phe Arg Gly Pro Pro Thr Gly Ile
145 150 155 160
Val Val Glu Arg Glu Arg Leu Asp Lys Phe Gly Arg Pro Leu Leu Gly
165 170 175
Ala Thr Thr Lys Pro Lys Leu Gly Leu Ser Gly Lys Asn Tyr Gly Arg
180 185 190
Val Val Tyr Glu Ala Leu Lys Gly Gly Leu Asp Phe Val Lys Asp Asp
195 200 205
Glu Asn Ile Asn Ser Gln Pro Phe Met His Trp Arg Asp Arg Phe Leu
210 215 220
Tyr Cys Met Glu Ala Val Asn Lys Ala Gln Ala Glu Thr Gly Glu Val
225 230 235 240
Lys Gly His Tyr Leu Asn Ile Thr Ala Gly Thr Met Glu Glu Met Tyr
245 250 255
Arg Arg Ala Glu Phe Ala Lys Glu Leu Gly Ser Val Val Val Met Val
260 265 270
Asp Leu Ile Ile Gly Trp Thr Ala Ile Gln Ser Met Ser Asn Trp Cys
275 280 285
Arg Glu Asn Asp Met Ile Leu His Met His Arg Ala Gly His Gly Thr
290 295 300
Tyr Thr Arg Gln Lys Ser His Gly Val Ser Phe Arg Val Ile Ala Lys
305 310 315 320
Trp Leu Arg Leu Ala Gly Val Asp His Leu His Thr Gly Thr Ala Val
325 330 335
Gly Lys Leu Glu Gly Asp Pro Met Thr Val Gln Gly Phe Tyr Asn Val
340 345 350
Cys Arg Glu Thr Thr Thr Gln Gln Asp Leu Thr Arg Gly Leu Phe Phe
355 360 365
Glu Gln Asp Trp Gly Gly Ile Arg Lys Val Met Pro Val Ala Ser Gly
370 375 380
Gly Ile His Ala Gly Gln Met His Gln Leu Ile Asp Leu Phe Gly Glu
385 390 395 400
Asp Val Val Leu Gln Phe Gly Gly Gly Thr Ile Gly His Pro Asp Gly
405 410 415
Ile Gln Ala Gly Ala Thr Ala Asn Arg Val Ala Leu Glu Thr Met Ile
420 425 430
Leu Ala Arg Asn Glu Gly Arg Asp Ile Arg Asn Glu Gly Pro Glu Ile
435 440 445
Leu Val Glu Ala Ala Lys Trp Cys Arg Pro Leu Arg Ala Ala Leu Asp
450 455 460
Thr Trp Gly Glu Val Thr Phe Asn Tyr Ala Ser Thr Asp Thr Ser Asp
465 470 475 480
Tyr Val Pro Thr Ala Ser Val Ala
485
<210> 4
<211> 405
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 4
atgcgcatca cccaaggctc cttctccttc ctgccggacc tcaccgacac gcagatcaag 60
gcccaggtgc aatattgcct ggaccagggc tgggcggtct cggtggagca caccgacgat 120
ccccacccgc gcaacaccta ttgggagatg tggggcccgc ccatgttcga tctgcgcgac 180
gcggccggcg tcttcggcga gatcgaagcc tgccgggccg ccaatcccga gcattatgtg 240
cgggtgaacg ccttcgattc cagccgcgga tgggagacga tccgcctgtc cttcatcgtt 300
cagcggccca ccgtggaaga gggcttccgc ctcgaccgca ccgaaggcaa gggccgcaac 360
cagagctacg ccatgcgcta ccgggcgcag ttcgcgccgc gctga 405
<210> 5
<211> 134
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 5
Met Arg Ile Thr Gln Gly Ser Phe Ser Phe Leu Pro Asp Leu Thr Asp
1 5 10 15
Thr Gln Ile Lys Ala Gln Val Gln Tyr Cys Leu Asp Gln Gly Trp Ala
20 25 30
Val Ser Val Glu His Thr Asp Asp Pro His Pro Arg Asn Thr Tyr Trp
35 40 45
Glu Met Trp Gly Pro Pro Met Phe Asp Leu Arg Asp Ala Ala Gly Val
50 55 60
Phe Gly Glu Ile Glu Ala Cys Arg Ala Ala Asn Pro Glu His Tyr Val
65 70 75 80
Arg Val Asn Ala Phe Asp Ser Ser Arg Gly Trp Glu Thr Ile Arg Leu
85 90 95
Ser Phe Ile Val Gln Arg Pro Thr Val Glu Glu Gly Phe Arg Leu Asp
100 105 110
Arg Thr Glu Gly Lys Gly Arg Asn Gln Ser Tyr Ala Met Arg Tyr Arg
115 120 125
Ala Gln Phe Ala Pro Arg
130
<210> 6
<211> 1896
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 6
atgatgccat ctgagccgca cggcgcgggc atgccgcccc cacgggaagc ggccgcggtt 60
cccacccccc aggaggtgag cgcggtggtg gccgaggtgg tcgcggatgc cgtggcatcg 120
gtgggcggcg cacgcacccg gctcatggac atcgtccagc tggcccagca gcgtctcggc 180
catctctccg aagagaccat ggcggccatt gccgcgcggc tcgccattcc gccggtggaa 240
gtggcggaca tggtgtcctt ctacgccttc ctgaaccgcg cgcccaaggg ccgctaccac 300
atccgcctgt cgcgcagccc catctcgctg atgaagggcg ccgaggcggt ggctgccgcc 360
ttctgccaga tcctcggcat cgccatgggc gagacctcgc aggatggcga cttcaccctg 420
gaatggacca acgacatcgg catggccgac caggagccgg ccgccctcgt caacggcacg 480
gtgatgacgc agctcgcgcc cggcgatgcg gccatcatcg tcggccggct gcgggcccat 540
cacgcgccca atgccctgcc gctgttccct ggagccggcg tggccggctc cggcctgccc 600
catgcccgga tccgccccag cctggtgatg ccgggacagc ttctgttccg cgaggaccac 660
acgacgccgg gcgccggcat caaggcggca ctcgccctca ccccggacga agtggtgcag 720
aaggtctccg ccgcgcgcct gcgcgggcgg ggtggcgccg gctttcccac cggtctcaaa 780
tggaagctct gccgccagtc gcccgccacc acccgccatg tgatctgcaa tgcggacgag 840
ggcgagcccg gcaccttcaa ggatcgcgtg ctgctcacgc aggcgccgca cctcatgttc 900
gacggcatga ccatcgccgg ctacgccttg ggggcgcggg agggcgtggt ctatctgcgc 960
ggcgagtacg cctatctgtg ggagcctctg catgcggtcc tgcgcgagcg ctatgggctc 1020
gggctcgccg gcgcgaacat cctgggacac gcgggcttcg acttcgacat ccgcatccag 1080
ctgggcgccg gcgcctatat ctgcggcgag gaatccgcgc tggtggaatc gctggaaggc 1140
aagcgcggct cgccccgcga ccgccccccc ttccccaccg tgcgcggcca tctccagcag 1200
cccaccgccg tggacaatgt ggagaccttc gcctgcgccg cccgcatcct ggaggatggc 1260
gtggaggcgt tcgcgggcat cggcacgccc gaatccgccg gcacgaagct cctctcggtg 1320
tcgggcgatt gcccgcgccc cggcgtgtat gaggtgccct tcggcctcac ggtgaacgcg 1380
ctgctcgacc ttgtcggcgc gccggacgcc gccttcgtgc agatgggtgg gccgtccggc 1440
caatgcgtgg cgccgaagga ttacggccgc cgcatcgcct tcgaggacct gcccaccggc 1500
ggctcggtga tggtgttcgg cccggggcgc gacgtgctcg ccatggtgcg cgagttcgcg 1560
gatttcttcg ccggcgaatc ctgcggctgg tgcacgccct gccgggtggg caccaccttg 1620
ctcaaggaag agctggacaa gctcctcgcc aaccgcgcca ccctcgccga catccgcgcg 1680
ctggagaccc tggccacgac cgtctcccgc accagccgct gcggcctcgg ccagacggcg 1740
cccaacccca tcctttccac catgcgcaac ctgccggaag cctatgaggc gaggctgagg 1800
cccgaagact tcctgccctg ggcctcgctc gacgaggcgc tgaagcccgc catcgtcatc 1860
cagggccgcg cgcccgtgcc ggaggaagag gcatga 1896
<210> 7
<211> 631
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 7
Met Met Pro Ser Glu Pro His Gly Ala Gly Met Pro Pro Pro Arg Glu
1 5 10 15
Ala Ala Ala Val Pro Thr Pro Gln Glu Val Ser Ala Val Val Ala Glu
20 25 30
Val Val Ala Asp Ala Val Ala Ser Val Gly Gly Ala Arg Thr Arg Leu
35 40 45
Met Asp Ile Val Gln Leu Ala Gln Gln Arg Leu Gly His Leu Ser Glu
50 55 60
Glu Thr Met Ala Ala Ile Ala Ala Arg Leu Ala Ile Pro Pro Val Glu
65 70 75 80
Val Ala Asp Met Val Ser Phe Tyr Ala Phe Leu Asn Arg Ala Pro Lys
85 90 95
Gly Arg Tyr His Ile Arg Leu Ser Arg Ser Pro Ile Ser Leu Met Lys
100 105 110
Gly Ala Glu Ala Val Ala Ala Ala Phe Cys Gln Ile Leu Gly Ile Ala
115 120 125
Met Gly Glu Thr Ser Gln Asp Gly Asp Phe Thr Leu Glu Trp Thr Asn
130 135 140
Asp Ile Gly Met Ala Asp Gln Glu Pro Ala Ala Leu Val Asn Gly Thr
145 150 155 160
Val Met Thr Gln Leu Ala Pro Gly Asp Ala Ala Ile Ile Val Gly Arg
165 170 175
Leu Arg Ala His His Ala Pro Asn Ala Leu Pro Leu Phe Pro Gly Ala
180 185 190
Gly Val Ala Gly Ser Gly Leu Pro His Ala Arg Ile Arg Pro Ser Leu
195 200 205
Val Met Pro Gly Gln Leu Leu Phe Arg Glu Asp His Thr Thr Pro Gly
210 215 220
Ala Gly Ile Lys Ala Ala Leu Ala Leu Thr Pro Asp Glu Val Val Gln
225 230 235 240
Lys Val Ser Ala Ala Arg Leu Arg Gly Arg Gly Gly Ala Gly Phe Pro
245 250 255
Thr Gly Leu Lys Trp Lys Leu Cys Arg Gln Ser Pro Ala Thr Thr Arg
260 265 270
His Val Ile Cys Asn Ala Asp Glu Gly Glu Pro Gly Thr Phe Lys Asp
275 280 285
Arg Val Leu Leu Thr Gln Ala Pro His Leu Met Phe Asp Gly Met Thr
290 295 300
Ile Ala Gly Tyr Ala Leu Gly Ala Arg Glu Gly Val Val Tyr Leu Arg
305 310 315 320
Gly Glu Tyr Ala Tyr Leu Trp Glu Pro Leu His Ala Val Leu Arg Glu
325 330 335
Arg Tyr Gly Leu Gly Leu Ala Gly Ala Asn Ile Leu Gly His Ala Gly
340 345 350
Phe Asp Phe Asp Ile Arg Ile Gln Leu Gly Ala Gly Ala Tyr Ile Cys
355 360 365
Gly Glu Glu Ser Ala Leu Val Glu Ser Leu Glu Gly Lys Arg Gly Ser
370 375 380
Pro Arg Asp Arg Pro Pro Phe Pro Thr Val Arg Gly His Leu Gln Gln
385 390 395 400
Pro Thr Ala Val Asp Asn Val Glu Thr Phe Ala Cys Ala Ala Arg Ile
405 410 415
Leu Glu Asp Gly Val Glu Ala Phe Ala Gly Ile Gly Thr Pro Glu Ser
420 425 430
Ala Gly Thr Lys Leu Leu Ser Val Ser Gly Asp Cys Pro Arg Pro Gly
435 440 445
Val Tyr Glu Val Pro Phe Gly Leu Thr Val Asn Ala Leu Leu Asp Leu
450 455 460
Val Gly Ala Pro Asp Ala Ala Phe Val Gln Met Gly Gly Pro Ser Gly
465 470 475 480
Gln Cys Val Ala Pro Lys Asp Tyr Gly Arg Arg Ile Ala Phe Glu Asp
485 490 495
Leu Pro Thr Gly Gly Ser Val Met Val Phe Gly Pro Gly Arg Asp Val
500 505 510
Leu Ala Met Val Arg Glu Phe Ala Asp Phe Phe Ala Gly Glu Ser Cys
515 520 525
Gly Trp Cys Thr Pro Cys Arg Val Gly Thr Thr Leu Leu Lys Glu Glu
530 535 540
Leu Asp Lys Leu Leu Ala Asn Arg Ala Thr Leu Ala Asp Ile Arg Ala
545 550 555 560
Leu Glu Thr Leu Ala Thr Thr Val Ser Arg Thr Ser Arg Cys Gly Leu
565 570 575
Gly Gln Thr Ala Pro Asn Pro Ile Leu Ser Thr Met Arg Asn Leu Pro
580 585 590
Glu Ala Tyr Glu Ala Arg Leu Arg Pro Glu Asp Phe Leu Pro Trp Ala
595 600 605
Ser Leu Asp Glu Ala Leu Lys Pro Ala Ile Val Ile Gln Gly Arg Ala
610 615 620
Pro Val Pro Glu Glu Glu Ala
625 630
<210> 8
<211> 1491
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 8
atgagccggg gatcccccga tgccgggaaa gaccgcacca tgagcgccac cgacggcacc 60
accgcccccc gcaagatcgt catcgatccg gtgacccgcg tggagggcca cggcaaggtc 120
accatccgcc tggatgaagc cggcgcggtg gaggatgcgc gtttccacat cgtggagttc 180
cgcggcttcg agcggttcat ccagggccgg atgtactggg aagtgcccct tatcatccag 240
cggctgtgcg gcatctgccc ggtgagccac catctggcgg cggcgaaagc catggaccag 300
gtggcgggcg tggaccgcgt accgcccacc gccgagaaac tgcgccggct gatgcattat 360
gggcaggtgc tgcaatccaa cgctttgcac atcttccacc tcgcctcgcc cgacctcctg 420
ttcggcttcg acgcgccggc cgagcagcgc aacatcatcg ccgtgctcca gcgttatccg 480
gagatcggca aatgggcgat cttcatcagg aagttcggcc aggaggtcat caaggccacc 540
ggcgggcgca agatccatcc caccagcgcc attcccggcg gggtcaacca gaacctcgcc 600
gtggaggacc gcgacgccct gcgcgccaag gtgggcgaga tcatcagctg gtgcatggcg 660
gcgctggacc atcacaaggc ctatgtggcg gaaaaccggg cgctgcatga cagcttcgcc 720
gccttcccct ccgccttcat gagcctcgtg gggccggatg gcggcatgga cctttatgac 780
ggcaccctgc gggtgatcga tgccgagggc gcccccctca tcgaaggcgc gccgcccgcc 840
tcctaccgcg accacctcat cgaggaggtg cggccctgga gctatctgaa attcccccat 900
ctgcgcgcct tcggccgcga cgatggctgg tatcgggtcg gccccctcgc ccaggtcaat 960
tgcgccgcgt ccatcgacac gccccgcgcc gaggcggccc ggcgggactt catggccgag 1020
ggcggcggca agccggtgca tgccaccctc gcttatcact gggcgcggct catcgtgctg 1080
gtccattgcg cggagaagat cgaacagctg ctgttcgacg acgacctgca aggctgcgat 1140
ctgcgtgcgg agggcacccg gcgcggggaa ggcgtcgcct ggatcgaggc gccgcgcggc 1200
accctcatcc accattacga ggtggacgag aacgaccagg tgcgccgcgc caacctcatc 1260
gtctccacca cccacaataa cgaggccatg aaccgcgccg tgcggcaggt ggcgaagacg 1320
gacctttccg gtcgcgagat caccgaaggg ctgctgaacc atatcgaggt ggccatccgc 1380
gccttcgacc cctgcctgtc ctgcgccacc catgcgctgg gccagatgcc gctgatcgtg 1440
acgcttgaag atgcctccgg cgcagagatc gcccgcggag tgaaggaatg a 1491
<210> 9
<211> 496
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 9
Met Ser Arg Gly Ser Pro Asp Ala Gly Lys Asp Arg Thr Met Ser Ala
1 5 10 15
Thr Asp Gly Thr Thr Ala Pro Arg Lys Ile Val Ile Asp Pro Val Thr
20 25 30
Arg Val Glu Gly His Gly Lys Val Thr Ile Arg Leu Asp Glu Ala Gly
35 40 45
Ala Val Glu Asp Ala Arg Phe His Ile Val Glu Phe Arg Gly Phe Glu
50 55 60
Arg Phe Ile Gln Gly Arg Met Tyr Trp Glu Val Pro Leu Ile Ile Gln
65 70 75 80
Arg Leu Cys Gly Ile Cys Pro Val Ser His His Leu Ala Ala Ala Lys
85 90 95
Ala Met Asp Gln Val Ala Gly Val Asp Arg Val Pro Pro Thr Ala Glu
100 105 110
Lys Leu Arg Arg Leu Met His Tyr Gly Gln Val Leu Gln Ser Asn Ala
115 120 125
Leu His Ile Phe His Leu Ala Ser Pro Asp Leu Leu Phe Gly Phe Asp
130 135 140
Ala Pro Ala Glu Gln Arg Asn Ile Ile Ala Val Leu Gln Arg Tyr Pro
145 150 155 160
Glu Ile Gly Lys Trp Ala Ile Phe Ile Arg Lys Phe Gly Gln Glu Val
165 170 175
Ile Lys Ala Thr Gly Gly Arg Lys Ile His Pro Thr Ser Ala Ile Pro
180 185 190
Gly Gly Val Asn Gln Asn Leu Ala Val Glu Asp Arg Asp Ala Leu Arg
195 200 205
Ala Lys Val Gly Glu Ile Ile Ser Trp Cys Met Ala Ala Leu Asp His
210 215 220
His Lys Ala Tyr Val Ala Glu Asn Arg Ala Leu His Asp Ser Phe Ala
225 230 235 240
Ala Phe Pro Ser Ala Phe Met Ser Leu Val Gly Pro Asp Gly Gly Met
245 250 255
Asp Leu Tyr Asp Gly Thr Leu Arg Val Ile Asp Ala Glu Gly Ala Pro
260 265 270
Leu Ile Glu Gly Ala Pro Pro Ala Ser Tyr Arg Asp His Leu Ile Glu
275 280 285
Glu Val Arg Pro Trp Ser Tyr Leu Lys Phe Pro His Leu Arg Ala Phe
290 295 300
Gly Arg Asp Asp Gly Trp Tyr Arg Val Gly Pro Leu Ala Gln Val Asn
305 310 315 320
Cys Ala Ala Ser Ile Asp Thr Pro Arg Ala Glu Ala Ala Arg Arg Asp
325 330 335
Phe Met Ala Glu Gly Gly Gly Lys Pro Val His Ala Thr Leu Ala Tyr
340 345 350
His Trp Ala Arg Leu Ile Val Leu Val His Cys Ala Glu Lys Ile Glu
355 360 365
Gln Leu Leu Phe Asp Asp Asp Leu Gln Gly Cys Asp Leu Arg Ala Glu
370 375 380
Gly Thr Arg Arg Gly Glu Gly Val Ala Trp Ile Glu Ala Pro Arg Gly
385 390 395 400
Thr Leu Ile His His Tyr Glu Val Asp Glu Asn Asp Gln Val Arg Arg
405 410 415
Ala Asn Leu Ile Val Ser Thr Thr His Asn Asn Glu Ala Met Asn Arg
420 425 430
Ala Val Arg Gln Val Ala Lys Thr Asp Leu Ser Gly Arg Glu Ile Thr
435 440 445
Glu Gly Leu Leu Asn His Ile Glu Val Ala Ile Arg Ala Phe Asp Pro
450 455 460
Cys Leu Ser Cys Ala Thr His Ala Leu Gly Gln Met Pro Leu Ile Val
465 470 475 480
Thr Leu Glu Asp Ala Ser Gly Ala Glu Ile Ala Arg Gly Val Lys Glu
485 490 495
<210> 10
<211> 720
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 10
atgagcgaga cccccttcac ctttaccgtg gacggcatcg cggtcccggc cacccccggc 60
cagagcgtca tcgaggcgtg cgatgcggcg ggcatctata tcccgcgcct gtgccaccac 120
ccggacctgc cgccggcggg ccattgccgg gtgtgcacct gcatcatcga cgggcggccg 180
gccagcgcct gcaccatgcc cgccgccagg ggcatggtgg tggagaacga gacgcccgct 240
ttgctggcgg agcggcgcac gctgatcgag atgctgttcg cggaaggcaa ccatttctgc 300
cagttctgcg aggcgagcgg cgattgcgaa ttgcaggcgc tgggctacct gttcggcatg 360
gtggccccgc ccttccccca tctgtggccg aagcggccgg tggatgccag ccatccggat 420
atctatatcg accacaatcg ctgcatcctg tgctcgcgct gcgtgcgcgc ctcgcgcacc 480
ctggacggca agtccgtgtt cggcttcgag gggcgcggca tcgagatgca tctggcggtg 540
accggcgggc acctggacga cagcgccatc gccgccgccg acagggcggt tgagatgtgc 600
ccggtgggct gcatcgtcct caagcgcacc ggctaccgca cgccctatgg ccggcggcgc 660
tacgacgccg cgcccatcgg ctccgacatc accgcccggc gcggcggcgc gaaggactga 720
<210> 11
<211> 239
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 11
Met Ser Glu Thr Pro Phe Thr Phe Thr Val Asp Gly Ile Ala Val Pro
1 5 10 15
Ala Thr Pro Gly Gln Ser Val Ile Glu Ala Cys Asp Ala Ala Gly Ile
20 25 30
Tyr Ile Pro Arg Leu Cys His His Pro Asp Leu Pro Pro Ala Gly His
35 40 45
Cys Arg Val Cys Thr Cys Ile Ile Asp Gly Arg Pro Ala Ser Ala Cys
50 55 60
Thr Met Pro Ala Ala Arg Gly Met Val Val Glu Asn Glu Thr Pro Ala
65 70 75 80
Leu Leu Ala Glu Arg Arg Thr Leu Ile Glu Met Leu Phe Ala Glu Gly
85 90 95
Asn His Phe Cys Gln Phe Cys Glu Ala Ser Gly Asp Cys Glu Leu Gln
100 105 110
Ala Leu Gly Tyr Leu Phe Gly Met Val Ala Pro Pro Phe Pro His Leu
115 120 125
Trp Pro Lys Arg Pro Val Asp Ala Ser His Pro Asp Ile Tyr Ile Asp
130 135 140
His Asn Arg Cys Ile Leu Cys Ser Arg Cys Val Arg Ala Ser Arg Thr
145 150 155 160
Leu Asp Gly Lys Ser Val Phe Gly Phe Glu Gly Arg Gly Ile Glu Met
165 170 175
His Leu Ala Val Thr Gly Gly His Leu Asp Asp Ser Ala Ile Ala Ala
180 185 190
Ala Asp Arg Ala Val Glu Met Cys Pro Val Gly Cys Ile Val Leu Lys
195 200 205
Arg Thr Gly Tyr Arg Thr Pro Tyr Gly Arg Arg Arg Tyr Asp Ala Ala
210 215 220
Pro Ile Gly Ser Asp Ile Thr Ala Arg Arg Gly Gly Ala Lys Asp
225 230 235
<210> 12
<211> 543
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 12
atggccaagc ccaaactcgc cacctgcgcg ctggccggct gcttcggctg ccacatgtcc 60
ttcctggaca tggacgagcg catcgtcgag ctcatcgacc tggtggacct cgacgtctcg 120
cccctcgacg acaagaaaaa cttcaccggc atggtggaaa tcggcctggt ggaaggcggc 180
tgcgccgacg agcgccatgt gaaggtgctg cgcgagttcc gcgagaaatc ccgcatcctg 240
gtggcggtgg gcgcctgcgc catcaccggc ggcatcccgg cattgcgcaa cctcgccggc 300
ctcgacgaat gcctgaggga agcctacctc accggcccca cggtggaagg cggcgggctc 360
attcccaacg acccggagct gccgctgctg ctggacaagg tctatccggt gcaggacttc 420
gtgaagatcg accatttcct gcccggctgc ccgccctcgg ccgacgccat ctgggcggct 480
ctgaaggcgc tgctgaccgg caccgagccg catctgccct acccgctttt caagtacgaa 540
tga 543
<210> 13
<211> 180
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 13
Met Ala Lys Pro Lys Leu Ala Thr Cys Ala Leu Ala Gly Cys Phe Gly
1 5 10 15
Cys His Met Ser Phe Leu Asp Met Asp Glu Arg Ile Val Glu Leu Ile
20 25 30
Asp Leu Val Asp Leu Asp Val Ser Pro Leu Asp Asp Lys Lys Asn Phe
35 40 45
Thr Gly Met Val Glu Ile Gly Leu Val Glu Gly Gly Cys Ala Asp Glu
50 55 60
Arg His Val Lys Val Leu Arg Glu Phe Arg Glu Lys Ser Arg Ile Leu
65 70 75 80
Val Ala Val Gly Ala Cys Ala Ile Thr Gly Gly Ile Pro Ala Leu Arg
85 90 95
Asn Leu Ala Gly Leu Asp Glu Cys Leu Arg Glu Ala Tyr Leu Thr Gly
100 105 110
Pro Thr Val Glu Gly Gly Gly Leu Ile Pro Asn Asp Pro Glu Leu Pro
115 120 125
Leu Leu Leu Asp Lys Val Tyr Pro Val Gln Asp Phe Val Lys Ile Asp
130 135 140
His Phe Leu Pro Gly Cys Pro Pro Ser Ala Asp Ala Ile Trp Ala Ala
145 150 155 160
Leu Lys Ala Leu Leu Thr Gly Thr Glu Pro His Leu Pro Tyr Pro Leu
165 170 175
Phe Lys Tyr Glu
180
<210> 14
<211> 1013
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 14
tccagacccg ggcaacattg ctccatgtgc tgggcaccct ggccggccgc tggccccata 60
ccctcgcgct ccagcccggc ggggtgaccc gaagcgccga ccagcacgac cgcatgcgcc 120
tgctcgcgac gctgaaggcg gtgcgggcgg cgctggaaga gaccttgttc ggcgcgcctt 180
tggaagaggt ggcggccctg gacggcgccg ccgccgtgga ggcctggcgc gccaacggcc 240
cggaagggga tttccgcctg ttcctggaga tcgccgccga cctggagctg gaccggctcg 300
gccgcgcgca cgaccgcttt ctctccttcg gcgcctacgc ccaggacgag gggcgccttt 360
atggcgccgg caccttcgag gccgggacgg cgggagggct cgatcccaac gccatcaccg 420
aggaccacgc cttcgcccgc atggaggacc gcgcggcgcc ccatgcgccc tttgacggct 480
ccaccttccc cgatgccgac gacaccgagg gctacacctg gtgcaaggcg ccgcgccttg 540
ccggcctgcc cttcgagacc ggcgccttcg cccggcaggt ggtggcgggc catccgctcg 600
cccgggacct cgtgacgcgg gaaggcggca ctgtgcgcag ccgcgtggtc ggccggctgc 660
tggaaaccgc gcgcaccctg atcgccatgg agggctgggt gaaggaactg cggcccgaag 720
ggccctggtg cgcccagggc cacctgcccc aggaaggccg cgccttcggc ctcaccgagg 780
cggcgcgcgg ggcgctcggc cactggatgg tggtggagaa gggccgcatt gcccgctacc 840
agatcatcgc ccccaccacc tggaacttct ccccccgcga cggcgcgggc ctgcccggcc 900
cgctggagac ggccctggtg ggcgcgcccg tgcggcaggg agagacgacg cccgtgagcg 960
tgcagcacat cgtgcgctcc ttcgacccgt gcatggtctg cactgtgcat tga 1013
<210> 15
<211> 485
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 15
Met Ser Ala Glu Thr Arg Arg Leu Val Val Gly Pro Phe Asn Arg Val
1 5 10 15
Glu Gly Asp Leu Glu Val Arg Leu Asp Val Gln Asp Gly Arg Val Gln
20 25 30
Gln Ala Phe Val Ser Ser Pro Leu Phe Arg Gly Phe Glu Arg Ile Leu
35 40 45
Glu Gly Arg Asp Pro Arg Asp Ala Leu Val Ile Ala Pro Arg Ile Cys
50 55 60
Gly Ile Cys Ser Val Ser Gln Ser His Ala Ala Ala Leu Ala Leu Ala
65 70 75 80
Gly Leu Gln Gly Ile Ala Pro Thr His Asp Gly Arg Ile Ala Thr Asn
85 90 95
Leu Ile Val Ala Ala Glu Asn Val Ala Asp His Leu Thr His Phe His
100 105 110
Val Phe Phe Met Pro Asp Phe Ala Arg Ala Val Tyr Glu Asp Arg Pro
115 120 125
Trp Phe Ala Gln Ala Ala Arg Arg Phe Lys Ala Asn Gln Gly Val Ser
130 135 140
Val Arg Arg Ala Leu Gln Thr Arg Ala Thr Leu Leu His Val Leu Gly
145 150 155 160
Thr Leu Ala Gly Arg Trp Pro His Thr Leu Ala Leu Gln Pro Gly Gly
165 170 175
Val Thr Arg Ser Ala Asp Gln His Asp Arg Met Arg Leu Leu Ala Thr
180 185 190
Leu Lys Ala Val Arg Ala Ala Leu Glu Glu Thr Leu Phe Gly Ala Pro
195 200 205
Leu Glu Glu Val Ala Ala Leu Asp Gly Ala Ala Ala Val Glu Ala Trp
210 215 220
Arg Ala Asn Gly Pro Glu Gly Asp Phe Arg Leu Phe Leu Glu Ile Ala
225 230 235 240
Ala Asp Leu Glu Leu Asp Arg Leu Gly Arg Ala His Asp Arg Phe Leu
245 250 255
Ser Phe Gly Ala Tyr Ala Gln Asp Glu Gly Arg Leu Tyr Gly Ala Gly
260 265 270
Thr Phe Glu Ala Gly Thr Ala Gly Gly Leu Asp Pro Asn Ala Ile Thr
275 280 285
Glu Asp His Ala Phe Ala Arg Met Glu Asp Arg Ala Ala Pro His Ala
290 295 300
Pro Phe Asp Gly Ser Thr Phe Pro Asp Ala Asp Asp Thr Glu Gly Tyr
305 310 315 320
Thr Trp Cys Lys Ala Pro Arg Leu Ala Gly Leu Pro Phe Glu Thr Gly
325 330 335
Ala Phe Ala Arg Gln Val Val Ala Gly His Pro Leu Ala Arg Asp Leu
340 345 350
Val Thr Arg Glu Gly Gly Thr Val Arg Ser Arg Val Val Gly Arg Leu
355 360 365
Leu Glu Thr Ala Arg Thr Leu Ile Ala Met Glu Gly Trp Val Lys Glu
370 375 380
Leu Arg Pro Glu Gly Pro Trp Cys Ala Gln Gly His Leu Pro Gln Glu
385 390 395 400
Gly Arg Ala Phe Gly Leu Thr Glu Ala Ala Arg Gly Ala Leu Gly His
405 410 415
Trp Met Val Val Glu Lys Gly Arg Ile Ala Arg Tyr Gln Ile Ile Ala
420 425 430
Pro Thr Thr Trp Asn Phe Ser Pro Arg Asp Gly Ala Gly Leu Pro Gly
435 440 445
Pro Leu Glu Thr Ala Leu Val Gly Ala Pro Val Arg Gln Gly Glu Thr
450 455 460
Thr Pro Val Ser Val Gln His Ile Val Arg Ser Phe Asp Pro Cys Met
465 470 475 480
Val Cys Thr Val His
485
<210> 16
<211> 858
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 16
acgggggagg aagcccgcgc catcttcgac gccatccttg ccggcgttat cgtcctcgac 60
gccctgtgcg tggaaggcgc gctgctgcgc gggccgaacg gcaccgggcg cttccatgtg 120
ctggcgggca cggacacccc caccatcgac tgggcgcggc agctcgccgg catggcgcgc 180
cacgtggtgg cggtgggcac ctgcgccgcc tatgggggcg tgacggcggc gggcatcaac 240
cccaccgatg cctgcggcct ccagttcgac ggacgccgga agggtggggc gctgggggcg 300
gacttccgct cccgctcggg gcttccggtc atcaatgtgg ccggctgccc cacccatccc 360
aactgggtga cggaaaccct gatgctgctc gcctgcggcc tgctgggcga ggccgacctc 420
gacgtctatg gccgcccgcg cttctatgcg gacctgctgg tgcatcacgg ctgcccgcgc 480
aacgaatact atgaatacaa ggcgagcgcc gagaagatga gcgacctcgg ctgcatgatg 540
gagcatctgg gctgcctcgg cacccaggcc cacgccgact gcaacacgcg cctttggaat 600
ggcgagggct cgtgcacccg cggcggctat gcctgcatca actgcacggc gccggaattc 660
gaggagccgg gccacgcctt cctggagacg cccaagatcg gcggcatccc catcggcctg 720
cccaccgaca tgcccaaggc ctggttcatc gccttgtcct ccctcgccaa ggcggcgacg 780
ccggagcggc tgcgcaagaa cgcggtgtcc gaccatgtgg tcacgccgcc cgccgtcaag 840
gacatcaagc ggcgatga 858
<210> 17
<211> 335
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 17
Met Ser Thr Pro Phe Ser Val Leu Trp Leu Gln Ser Gly Gly Cys Gly
1 5 10 15
Gly Cys Thr Met Ser Leu Leu Cys Ala Glu Ala Pro Asp Leu Ala Thr
20 25 30
Thr Leu Asp Ala Ala Gly Ile Gly Phe Leu Trp His Pro Ala Leu Ser
35 40 45
Glu Glu Thr Gly Glu Glu Ala Arg Ala Ile Phe Asp Ala Ile Leu Ala
50 55 60
Gly Val Ile Val Leu Asp Ala Leu Cys Val Glu Gly Ala Leu Leu Arg
65 70 75 80
Gly Pro Asn Gly Thr Gly Arg Phe His Val Leu Ala Gly Thr Asp Thr
85 90 95
Pro Thr Ile Asp Trp Ala Arg Gln Leu Ala Gly Met Ala Arg His Val
100 105 110
Val Ala Val Gly Thr Cys Ala Ala Tyr Gly Gly Val Thr Ala Ala Gly
115 120 125
Ile Asn Pro Thr Asp Ala Cys Gly Leu Gln Phe Asp Gly Arg Arg Lys
130 135 140
Gly Gly Ala Leu Gly Ala Asp Phe Arg Ser Arg Ser Gly Leu Pro Val
145 150 155 160
Ile Asn Val Ala Gly Cys Pro Thr His Pro Asn Trp Val Thr Glu Thr
165 170 175
Leu Met Leu Leu Ala Cys Gly Leu Leu Gly Glu Ala Asp Leu Asp Val
180 185 190
Tyr Gly Arg Pro Arg Phe Tyr Ala Asp Leu Leu Val His His Gly Cys
195 200 205
Pro Arg Asn Glu Tyr Tyr Glu Tyr Lys Ala Ser Ala Glu Lys Met Ser
210 215 220
Asp Leu Gly Cys Met Met Glu His Leu Gly Cys Leu Gly Thr Gln Ala
225 230 235 240
His Ala Asp Cys Asn Thr Arg Leu Trp Asn Gly Glu Gly Ser Cys Thr
245 250 255
Arg Gly Gly Tyr Ala Cys Ile Asn Cys Thr Ala Pro Glu Phe Glu Glu
260 265 270
Pro Gly His Ala Phe Leu Glu Thr Pro Lys Ile Gly Gly Ile Pro Ile
275 280 285
Gly Leu Pro Thr Asp Met Pro Lys Ala Trp Phe Ile Ala Leu Ser Ser
290 295 300
Leu Ala Lys Ala Ala Thr Pro Glu Arg Leu Arg Lys Asn Ala Val Ser
305 310 315 320
Asp His Val Val Thr Pro Pro Ala Val Lys Asp Ile Lys Arg Arg
325 330 335
<210> 18
<211> 870
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 18
gtgaccgagc gcctgtccga cgtcaacgcc cgcatcgcct cggtgcggca gctctcatcg 60
gtcatcacgg ccatgcgggg cattgcggcg gcgcgggcgc gggaggcgcg gggtcggctc 120
gacggcatcc gcgcctatgc gcagaccatc gccgaggcca tcggccatgt gctcgccgtg 180
ctgcccgagg aggcccgcgc ccggtcctcc gggcaccggc atcggggcca tgcggtcatc 240
gccctgtgcg cggagcaggg ctttgccggc gtcttcaacg agcgggtgct ggacgaggcc 300
gcccggctgc tgaccggcgg ggcggggccg gccgagctgc tgctggtggg cgaccggggc 360
ctgatggtgg cccgcgagcg ggggctcgat gtctcctggt cggtgcccat ggtggcccat 420
gcgggccagg cctcggcgct ggcggaccgc atcagcgagg agctctaccg gcggatcgat 480
gcgggacggg tgacgcgggt gtcggtggtg cacgccgagc ccgccgcgtc cgccgccatc 540
gagacggtgg tgaaagtgct ggtgccgttc gacttcgccc gcttccccct ggcgcgggtg 600
gcatccgccc cgctcatgac catgccgccg ccgcggctgc tggcccagct gtcggaggaa 660
tatgtgttcg ccgagctgtg cgaggcgctc accttgtcct tcgcggcgga gaacgaggcc 720
cgcatgcggg ccatgatcgc cgcccgcgcc aatgtggccg ataccctgga gggcctcgtc 780
ggccgcgccc ggcagatgcg ccaggaggag atcaccaacg agatcatcga gctggaaggc 840
ggcgccggca gcgcccggca tgcggattga 870
<210> 19
<211> 289
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 19
Met Thr Glu Arg Leu Ser Asp Val Asn Ala Arg Ile Ala Ser Val Arg
1 5 10 15
Gln Leu Ser Ser Val Ile Thr Ala Met Arg Gly Ile Ala Ala Ala Arg
20 25 30
Ala Arg Glu Ala Arg Gly Arg Leu Asp Gly Ile Arg Ala Tyr Ala Gln
35 40 45
Thr Ile Ala Glu Ala Ile Gly His Val Leu Ala Val Leu Pro Glu Glu
50 55 60
Ala Arg Ala Arg Ser Ser Gly His Arg His Arg Gly His Ala Val Ile
65 70 75 80
Ala Leu Cys Ala Glu Gln Gly Phe Ala Gly Val Phe Asn Glu Arg Val
85 90 95
Leu Asp Glu Ala Ala Arg Leu Leu Thr Gly Gly Ala Gly Pro Ala Glu
100 105 110
Leu Leu Leu Val Gly Asp Arg Gly Leu Met Val Ala Arg Glu Arg Gly
115 120 125
Leu Asp Val Ser Trp Ser Val Pro Met Val Ala His Ala Gly Gln Ala
130 135 140
Ser Ala Leu Ala Asp Arg Ile Ser Glu Glu Leu Tyr Arg Arg Ile Asp
145 150 155 160
Ala Gly Arg Val Thr Arg Val Ser Val Val His Ala Glu Pro Ala Ala
165 170 175
Ser Ala Ala Ile Glu Thr Val Val Lys Val Leu Val Pro Phe Asp Phe
180 185 190
Ala Arg Phe Pro Leu Ala Arg Val Ala Ser Ala Pro Leu Met Thr Met
195 200 205
Pro Pro Pro Arg Leu Leu Ala Gln Leu Ser Glu Glu Tyr Val Phe Ala
210 215 220
Glu Leu Cys Glu Ala Leu Thr Leu Ser Phe Ala Ala Glu Asn Glu Ala
225 230 235 240
Arg Met Arg Ala Met Ile Ala Ala Arg Ala Asn Val Ala Asp Thr Leu
245 250 255
Glu Gly Leu Val Gly Arg Ala Arg Gln Met Arg Gln Glu Glu Ile Thr
260 265 270
Asn Glu Ile Ile Glu Leu Glu Gly Gly Ala Gly Ser Ala Arg His Ala
275 280 285
Asp
<210> 20
<211> 1536
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 20
atgagcacgg gcgcgcaagc gagcgaggat tggctcaccc ggagccgggc ggccctggcc 60
gggacgcgcc tttcccagca atcccaatcg gtgggccggg tggaggagat ggccgacggc 120
atcgcccgcg tctccggcct gccggatgtg cggctcgacg agcttctcac cttcgagggc 180
ggccagaccg gctatgccct caccctcgat cgcaccgaga tcgccgtggt gctgctggat 240
gacgcctccg gcgtggaggc gggcgcccgg gtgttcggca ccggcgaggt ggtgaaggtg 300
ccggtggggc cggggctgct gggccgcatc gtcgaccccc tcggccggcc catggaccgc 360
tccgagccgg tggtggcgca ggcgcaccat cccatcgagc ggccggcgcc ggccatcatc 420
gcccgcgacc tggtctcgca gccggttcag accggcacgc tggtggtgga tgcgctgttc 480
tccctcggcc ggggccagcg cgagctcatc atcggcgacc gggctaccgg caagaccgcc 540
atcgcggtgg acaccatcat cagccagaag cattcggaca tcgtgtgcat ctacgtggcg 600
gtgggccagc gcgccgccgc cgtggagcgg gtggtggagg cggtgcgcgc ccacggggcg 660
atcgagcgct gcatcttcgt ggtcgcctcg gccgccgcct cgccagggct gcaatggatc 720
gcgccgttcg ccggcatgac catggcggaa tatttccgcg acaacggcca gcatgcgctc 780
atcatcatcg atgatctcac caagcatgcg gccacccatc gcgagctggc gctgctcacc 840
cacgagccgc cgggccgcga ggcctatccc ggcgacatct tctatgtgca cgcccgcctt 900
ctggagcggg ccgccaagct ctccgccgag ctgggcggtg gctcgctcac ggccctgccc 960
atcgcggaga cggacgcggg aaacctctcc gcctatatcc ccaccaacct catctccatc 1020
accgatgggc agatcgtgct ggattcgcgg ctgttcgcgg ccaaccagcg cccggcggtg 1080
gatgtgggcc tctccgtgag ccgggtgggc ggcaaggcgc agcatcccgc gcttcgggcc 1140
gtgtccgggc gcatccggct cgattattcc cagttcctgg agctggaaat gttcacccgc 1200
ttcggcggca tcaccgatac ccgcgtgaag gcgcagatca cccggggcga gcgcatccgc 1260
gcgctgctca cccagccgcg cttttccacc ctgcgccttc aggacgaggt ggcgctgctg 1320
gccgcgctgg cggagggggt gttcgacact ttggccccgg ggctgatggg cgccgtgcgt 1380
gcccgcattc cggcccagct ggatgcgcag gtgaaggacg tggcctcggc cctcgccgag 1440
ggcaaggtgc tggaggaggg cttgcacgcc cgtctcgtgg cggccgtgcg ggccgtcgcg 1500
gcggacgtgg ccgcgaccgc gaaggccggg ccgtga 1536
<210> 21
<211> 511
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 21
Met Ser Thr Gly Ala Gln Ala Ser Glu Asp Trp Leu Thr Arg Ser Arg
1 5 10 15
Ala Ala Leu Ala Gly Thr Arg Leu Ser Gln Gln Ser Gln Ser Val Gly
20 25 30
Arg Val Glu Glu Met Ala Asp Gly Ile Ala Arg Val Ser Gly Leu Pro
35 40 45
Asp Val Arg Leu Asp Glu Leu Leu Thr Phe Glu Gly Gly Gln Thr Gly
50 55 60
Tyr Ala Leu Thr Leu Asp Arg Thr Glu Ile Ala Val Val Leu Leu Asp
65 70 75 80
Asp Ala Ser Gly Val Glu Ala Gly Ala Arg Val Phe Gly Thr Gly Glu
85 90 95
Val Val Lys Val Pro Val Gly Pro Gly Leu Leu Gly Arg Ile Val Asp
100 105 110
Pro Leu Gly Arg Pro Met Asp Arg Ser Glu Pro Val Val Ala Gln Ala
115 120 125
His His Pro Ile Glu Arg Pro Ala Pro Ala Ile Ile Ala Arg Asp Leu
130 135 140
Val Ser Gln Pro Val Gln Thr Gly Thr Leu Val Val Asp Ala Leu Phe
145 150 155 160
Ser Leu Gly Arg Gly Gln Arg Glu Leu Ile Ile Gly Asp Arg Ala Thr
165 170 175
Gly Lys Thr Ala Ile Ala Val Asp Thr Ile Ile Ser Gln Lys His Ser
180 185 190
Asp Ile Val Cys Ile Tyr Val Ala Val Gly Gln Arg Ala Ala Ala Val
195 200 205
Glu Arg Val Val Glu Ala Val Arg Ala His Gly Ala Ile Glu Arg Cys
210 215 220
Ile Phe Val Val Ala Ser Ala Ala Ala Ser Pro Gly Leu Gln Trp Ile
225 230 235 240
Ala Pro Phe Ala Gly Met Thr Met Ala Glu Tyr Phe Arg Asp Asn Gly
245 250 255
Gln His Ala Leu Ile Ile Ile Asp Asp Leu Thr Lys His Ala Ala Thr
260 265 270
His Arg Glu Leu Ala Leu Leu Thr His Glu Pro Pro Gly Arg Glu Ala
275 280 285
Tyr Pro Gly Asp Ile Phe Tyr Val His Ala Arg Leu Leu Glu Arg Ala
290 295 300
Ala Lys Leu Ser Ala Glu Leu Gly Gly Gly Ser Leu Thr Ala Leu Pro
305 310 315 320
Ile Ala Glu Thr Asp Ala Gly Asn Leu Ser Ala Tyr Ile Pro Thr Asn
325 330 335
Leu Ile Ser Ile Thr Asp Gly Gln Ile Val Leu Asp Ser Arg Leu Phe
340 345 350
Ala Ala Asn Gln Arg Pro Ala Val Asp Val Gly Leu Ser Val Ser Arg
355 360 365
Val Gly Gly Lys Ala Gln His Pro Ala Leu Arg Ala Val Ser Gly Arg
370 375 380
Ile Arg Leu Asp Tyr Ser Gln Phe Leu Glu Leu Glu Met Phe Thr Arg
385 390 395 400
Phe Gly Gly Ile Thr Asp Thr Arg Val Lys Ala Gln Ile Thr Arg Gly
405 410 415
Glu Arg Ile Arg Ala Leu Leu Thr Gln Pro Arg Phe Ser Thr Leu Arg
420 425 430
Leu Gln Asp Glu Val Ala Leu Leu Ala Ala Leu Ala Glu Gly Val Phe
435 440 445
Asp Thr Leu Ala Pro Gly Leu Met Gly Ala Val Arg Ala Arg Ile Pro
450 455 460
Ala Gln Leu Asp Ala Gln Val Lys Asp Val Ala Ser Ala Leu Ala Glu
465 470 475 480
Gly Lys Val Leu Glu Glu Gly Leu His Ala Arg Leu Val Ala Ala Val
485 490 495
Arg Ala Val Ala Ala Asp Val Ala Ala Thr Ala Lys Ala Gly Pro
500 505 510
<210> 22
<211> 741
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 22
atgcagatcg actggtggac gctgggcctg cagacggtca acgtcctcgt tctcatctgg 60
ctcctgagcc gcttcctgtt caagccggtg gcgcaggtca tcgcgcagcg ccgtgccgag 120
atcgagaagc tggtggagga tgcgcgcgcc gccaaggccg ccgccgaggc cgagcgggac 180
acggcgaagg cggaggaggc gcgccttgcc gccgagcgcg gcgcccgcat ggcggcggtc 240
gccaaggagg cggaggcgca gaaggcggca ttgctggccg ccgccaagac cgaggccgag 300
gccctgcacg cggccgcgga agcggccatc gtccgggcgc gggcgagcga ggaggaagcc 360
gccgccgacc gcgccagccg ccttgccgtg gacatcgccg ccaagctgct ggaccggctg 420
cccgacgacg cccgggtcgc gggcttcatc gatggcctcg ccgaggggct tgaagccctg 480
cccgaggcga gccgggcggt gatcggcgtc gacggcgcgc cagtgcgcgt gacggccgcg 540
cgcgccctta tgccggcgga ggaggaggcc tgccgcacgc ggctctccca ggcgctgggc 600
cgtccggtga cgctggccgt gaccatcgac cccgccctca tcgccggcct ggagatggag 660
acgccccacg cggtggtgcg caattccttc aaggccgatc tcgaccgcgt caccgcggcg 720
ctcacccatc atgggacctg a 741
<210> 23
<211> 246
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 23
Met Gln Ile Asp Trp Trp Thr Leu Gly Leu Gln Thr Val Asn Val Leu
1 5 10 15
Val Leu Ile Trp Leu Leu Ser Arg Phe Leu Phe Lys Pro Val Ala Gln
20 25 30
Val Ile Ala Gln Arg Arg Ala Glu Ile Glu Lys Leu Val Glu Asp Ala
35 40 45
Arg Ala Ala Lys Ala Ala Ala Glu Ala Glu Arg Asp Thr Ala Lys Ala
50 55 60
Glu Glu Ala Arg Leu Ala Ala Glu Arg Gly Ala Arg Met Ala Ala Val
65 70 75 80
Ala Lys Glu Ala Glu Ala Gln Lys Ala Ala Leu Leu Ala Ala Ala Lys
85 90 95
Thr Glu Ala Glu Ala Leu His Ala Ala Ala Glu Ala Ala Ile Val Arg
100 105 110
Ala Arg Ala Ser Glu Glu Glu Ala Ala Ala Asp Arg Ala Ser Arg Leu
115 120 125
Ala Val Asp Ile Ala Ala Lys Leu Leu Asp Arg Leu Pro Asp Asp Ala
130 135 140
Arg Val Ala Gly Phe Ile Asp Gly Leu Ala Glu Gly Leu Glu Ala Leu
145 150 155 160
Pro Glu Ala Ser Arg Ala Val Ile Gly Val Asp Gly Ala Pro Val Arg
165 170 175
Val Thr Ala Ala Arg Ala Leu Met Pro Ala Glu Glu Glu Ala Cys Arg
180 185 190
Thr Arg Leu Ser Gln Ala Leu Gly Arg Pro Val Thr Leu Ala Val Thr
195 200 205
Ile Asp Pro Ala Leu Ile Ala Gly Leu Glu Met Glu Thr Pro His Ala
210 215 220
Val Val Arg Asn Ser Phe Lys Ala Asp Leu Asp Arg Val Thr Ala Ala
225 230 235 240
Leu Thr His His Gly Thr
245
<210> 24
<211> 243
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 24
atgactgtcg agatggtcag catcttcgcg gcggcgctcg ccgtctcctt cggcgccatc 60
gggccggccc tgggcgaggg ccgggcggtg gccgcggcca tggacgccat cgcccgccag 120
ccggaggcgg ccggaacctt gtcgcgcacg ctcttcgtcg gcctcgccat gatcgagacc 180
atggcgatct actgcctggt gatcgcgctc ctggtgctct tcgccaatcc gttcgtgaag 240
tga 243
<210> 25
<211> 80
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 25
Met Thr Val Glu Met Val Ser Ile Phe Ala Ala Ala Leu Ala Val Ser
1 5 10 15
Phe Gly Ala Ile Gly Pro Ala Leu Gly Glu Gly Arg Ala Val Ala Ala
20 25 30
Ala Met Asp Ala Ile Ala Arg Gln Pro Glu Ala Ala Gly Thr Leu Ser
35 40 45
Arg Thr Leu Phe Val Gly Leu Ala Met Ile Glu Thr Met Ala Ile Tyr
50 55 60
Cys Leu Val Ile Ala Leu Leu Val Leu Phe Ala Asn Pro Phe Val Lys
65 70 75 80
<210> 26
<211> 699
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 26
atgggctcgc cgctgatcct cgaacccctg ttccatatcg ggcccgtgcc catcaccgcg 60
ccggtggtgg tcacctggct catcatggcc gccttcattg ggctggcgcg gctcatcacc 120
cggaagcttt ccaccgatcc cacccggacc caggcggcgg tggaaacggt gctgaccgcc 180
atcgattccc agatcgccga caccatgcag gccgatcccg cgccttatcg cgcgctcatc 240
ggcaccatct tcctttatgt gctggtggcc aactggtcct cgctcatccc gggcatcgag 300
ccgcccacgg cgcatatcga gaccgatgcg gcgctcgctt tcatcgtgtt cgccgccacc 360
atcgggttcg ggttgaagac aaggggtgtg aagggctatc tcgccacctt cgccgaaccc 420
tcctgggtga tgatcccgct caatgtggtg gagcagatca cccggacctt ctcgctcatc 480
gtgcgcctgt tcggcaacat catgagcggg gtgttcgtgg tcggcatcat cctgtccctc 540
gccgggctgc tggtgcccat ccccctcatg gcgctcgatc tcctgaccgg cgccgtgcag 600
gcctacatct tcgcggtgct ggcctgcgtg ttcatcggcg cggccattgg cgaggcgccg 660
gcaaagcccc aatcgaagga gccagggaaa acatcatga 699
<210> 27
<211> 232
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 27
Met Gly Ser Pro Leu Ile Leu Glu Pro Leu Phe His Ile Gly Pro Val
1 5 10 15
Pro Ile Thr Ala Pro Val Val Val Thr Trp Leu Ile Met Ala Ala Phe
20 25 30
Ile Gly Leu Ala Arg Leu Ile Thr Arg Lys Leu Ser Thr Asp Pro Thr
35 40 45
Arg Thr Gln Ala Ala Val Glu Thr Val Leu Thr Ala Ile Asp Ser Gln
50 55 60
Ile Ala Asp Thr Met Gln Ala Asp Pro Ala Pro Tyr Arg Ala Leu Ile
65 70 75 80
Gly Thr Ile Phe Leu Tyr Val Leu Val Ala Asn Trp Ser Ser Leu Ile
85 90 95
Pro Gly Ile Glu Pro Pro Thr Ala His Ile Glu Thr Asp Ala Ala Leu
100 105 110
Ala Phe Ile Val Phe Ala Ala Thr Ile Gly Phe Gly Leu Lys Thr Arg
115 120 125
Gly Val Lys Gly Tyr Leu Ala Thr Phe Ala Glu Pro Ser Trp Val Met
130 135 140
Ile Pro Leu Asn Val Val Glu Gln Ile Thr Arg Thr Phe Ser Leu Ile
145 150 155 160
Val Arg Leu Phe Gly Asn Ile Met Ser Gly Val Phe Val Val Gly Ile
165 170 175
Ile Leu Ser Leu Ala Gly Leu Leu Val Pro Ile Pro Leu Met Ala Leu
180 185 190
Asp Leu Leu Thr Gly Ala Val Gln Ala Tyr Ile Phe Ala Val Leu Ala
195 200 205
Cys Val Phe Ile Gly Ala Ala Ile Gly Glu Ala Pro Ala Lys Pro Gln
210 215 220
Ser Lys Glu Pro Gly Lys Thr Ser
225 230
<210> 28
<211> 456
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 28
gtgagcgcgc cgctgcacct caccatcacc acgccggccg ccgttctggt ggaccgtgcc 60
gacatcgtgg ccctgcgtgc cgaggacgag agcggcagct tcggcatcct gcccggccat 120
gcggatttcc tgaccgttct ggaggcctgc gtggtgcgct tcaaggatgg ggccgacggc 180
gtgcattatt gtgctctcag tggtggcgtg ctgtcggtcg aggagggccg gcgcatcgcc 240
atcgcctgcc gtcagggcac ggtgagcgac gacctggtcg ccctggaagg ggcggtggac 300
gccatgcgtt cggcggagag cgatgccgac aagcgggccc gggtggagca gatgcgcctt 360
catgcccacg ccgtgcgcca gctcctgcac tatctgcggc ccggccgggc cggcggcgtg 420
gcgccggccg ccgcgccgga ggaggggccg tcatga 456
<210> 29
<211> 151
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 29
Met Ser Ala Pro Leu His Leu Thr Ile Thr Thr Pro Ala Ala Val Leu
1 5 10 15
Val Asp Arg Ala Asp Ile Val Ala Leu Arg Ala Glu Asp Glu Ser Gly
20 25 30
Ser Phe Gly Ile Leu Pro Gly His Ala Asp Phe Leu Thr Val Leu Glu
35 40 45
Ala Cys Val Val Arg Phe Lys Asp Gly Ala Asp Gly Val His Tyr Cys
50 55 60
Ala Leu Ser Gly Gly Val Leu Ser Val Glu Glu Gly Arg Arg Ile Ala
65 70 75 80
Ile Ala Cys Arg Gln Gly Thr Val Ser Asp Asp Leu Val Ala Leu Glu
85 90 95
Gly Ala Val Asp Ala Met Arg Ser Ala Glu Ser Asp Ala Asp Lys Arg
100 105 110
Ala Arg Val Glu Gln Met Arg Leu His Ala His Ala Val Arg Gln Leu
115 120 125
Leu His Tyr Leu Arg Pro Gly Arg Ala Gly Gly Val Ala Pro Ala Ala
130 135 140
Ala Pro Glu Glu Gly Pro Ser
145 150
<210> 30
<211> 1494
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 30
atggcagcgg cagatgagga ggcgcaatcg gccgccggcc ccgcctcggg ccgggtggtg 60
gccgtgcgcg gcgcggtgat cgacatcgcc tttgcccagc ctccgctgcc gccgctggac 120
gacgcccttc tcatcaccga cggccggggc ggcacggtgc tggtggaggt gcagagccat 180
atggatcggc acacggtgcg cgccatcgcc cttcaggcca ccaccggcct cagccggggg 240
ctggaggcgg cgcgggtggg cgggccggtg aaggtgccgg tgggagacca tgtgctcggc 300
cgcctcctgg atgtcaccgg cgccatcggc gacaagggcg ggccgctgcc ggccgacgtg 360
cccacgcggc cgatccacca cgcgccgcca tccttcgccg cgcagggcgg cacgtccgat 420
ctgtttcgca ccggcatcaa ggtcatcgac ctcctggcgc ccctcgccca gggcggcaag 480
gcggccatgt tcggcggggc cggcgtgggc aagaccgtgc tggtgatgga gctgatccac 540
gccatggtgg cgagctacaa gggcatctcg gtgtttgccg gcgtggggga gcgctcccgc 600
gagggccacg agatgctgct ggacatgacc gattccggcg tgctcgaccg caccgttctg 660
gtctatggcc agatgaacga gccccccggg gcccgctggc gggtgcccat gacggcgctg 720
accatcgccg aatatttccg cgacgagaag caccagaacg tcctgctgct gatggacaac 780
atcttccgct tcgtccaggc gggggcggag gtctccggcc ttttgggccg tccgccctcc 840
cgggtgggat accagccgac gctggcgagc gaggtggcgg cgctccagga acgcatcacc 900
tccgtgggcg aggcctcggt gaccgccatc gaggcggtct acgtgccggc ggatgacttc 960
accgatcccg ccgtgaccac catcgccgcc cacgtggatt ccatggtggt gctctcccgc 1020
gccatggcgg cggagggcat gtatccggcg gtggacccca tctcctcctc gtcggtgctg 1080
ctcgacccgc tcatcgtggg ggacgagcat gcgcgcgtcg ccaacgaggt gcgccggacc 1140
atcgagcatt atcgcgagct tcaggatgtg atctcgctgc tgggcatgga ggaattgggc 1200
accgaggatc gccgcatcgt ggagcgggcg cgccggctcc agcgcttcct cacccagccc 1260
ttcacggtca ccgaggcctt caccggcgtg cccggccgct cggtggccat cgccgacacc 1320
atcgccggct gcaggatgat cctgtccggc gcctgcgacg actggcagga aagcgccctc 1380
tacatggtgg gcaccatcga cgaggcccgc cagaaggagg aggccgctcg cgccaaggcg 1440
gggcagggcg ccccggccgg gacggcagcc gagacggcgg aggccgcccc gtga 1494
<210> 31
<211> 497
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 31
Met Ala Ala Ala Asp Glu Glu Ala Gln Ser Ala Ala Gly Pro Ala Ser
1 5 10 15
Gly Arg Val Val Ala Val Arg Gly Ala Val Ile Asp Ile Ala Phe Ala
20 25 30
Gln Pro Pro Leu Pro Pro Leu Asp Asp Ala Leu Leu Ile Thr Asp Gly
35 40 45
Arg Gly Gly Thr Val Leu Val Glu Val Gln Ser His Met Asp Arg His
50 55 60
Thr Val Arg Ala Ile Ala Leu Gln Ala Thr Thr Gly Leu Ser Arg Gly
65 70 75 80
Leu Glu Ala Ala Arg Val Gly Gly Pro Val Lys Val Pro Val Gly Asp
85 90 95
His Val Leu Gly Arg Leu Leu Asp Val Thr Gly Ala Ile Gly Asp Lys
100 105 110
Gly Gly Pro Leu Pro Ala Asp Val Pro Thr Arg Pro Ile His His Ala
115 120 125
Pro Pro Ser Phe Ala Ala Gln Gly Gly Thr Ser Asp Leu Phe Arg Thr
130 135 140
Gly Ile Lys Val Ile Asp Leu Leu Ala Pro Leu Ala Gln Gly Gly Lys
145 150 155 160
Ala Ala Met Phe Gly Gly Ala Gly Val Gly Lys Thr Val Leu Val Met
165 170 175
Glu Leu Ile His Ala Met Val Ala Ser Tyr Lys Gly Ile Ser Val Phe
180 185 190
Ala Gly Val Gly Glu Arg Ser Arg Glu Gly His Glu Met Leu Leu Asp
195 200 205
Met Thr Asp Ser Gly Val Leu Asp Arg Thr Val Leu Val Tyr Gly Gln
210 215 220
Met Asn Glu Pro Pro Gly Ala Arg Trp Arg Val Pro Met Thr Ala Leu
225 230 235 240
Thr Ile Ala Glu Tyr Phe Arg Asp Glu Lys His Gln Asn Val Leu Leu
245 250 255
Leu Met Asp Asn Ile Phe Arg Phe Val Gln Ala Gly Ala Glu Val Ser
260 265 270
Gly Leu Leu Gly Arg Pro Pro Ser Arg Val Gly Tyr Gln Pro Thr Leu
275 280 285
Ala Ser Glu Val Ala Ala Leu Gln Glu Arg Ile Thr Ser Val Gly Glu
290 295 300
Ala Ser Val Thr Ala Ile Glu Ala Val Tyr Val Pro Ala Asp Asp Phe
305 310 315 320
Thr Asp Pro Ala Val Thr Thr Ile Ala Ala His Val Asp Ser Met Val
325 330 335
Val Leu Ser Arg Ala Met Ala Ala Glu Gly Met Tyr Pro Ala Val Asp
340 345 350
Pro Ile Ser Ser Ser Ser Val Leu Leu Asp Pro Leu Ile Val Gly Asp
355 360 365
Glu His Ala Arg Val Ala Asn Glu Val Arg Arg Thr Ile Glu His Tyr
370 375 380
Arg Glu Leu Gln Asp Val Ile Ser Leu Leu Gly Met Glu Glu Leu Gly
385 390 395 400
Thr Glu Asp Arg Arg Ile Val Glu Arg Ala Arg Arg Leu Gln Arg Phe
405 410 415
Leu Thr Gln Pro Phe Thr Val Thr Glu Ala Phe Thr Gly Val Pro Gly
420 425 430
Arg Ser Val Ala Ile Ala Asp Thr Ile Ala Gly Cys Arg Met Ile Leu
435 440 445
Ser Gly Ala Cys Asp Asp Trp Gln Glu Ser Ala Leu Tyr Met Val Gly
450 455 460
Thr Ile Asp Glu Ala Arg Gln Lys Glu Glu Ala Ala Arg Ala Lys Ala
465 470 475 480
Gly Gln Gly Ala Pro Ala Gly Thr Ala Ala Glu Thr Ala Glu Ala Ala
485 490 495
Pro
<210> 32
<211> 1437
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 32
atggcgaaca aggtcggacg catcacccag atcatcggcg ccgtcgtcga cgtgcagttc 60
gacgggcatc tgccggcgat tctcaacgcg atcgagacca ccaaccaggg caaccggctg 120
gtgctcgaag tggctcagca tctcggcgag aacaccgtgc gctgcatcgc catggatgcc 180
actgaaggcc tggtgcgtgg ccaggaggtg gccgacaccg atgcgcccat ccaggtgccc 240
gtgggcgccg ccaccctcgg ccgcatcatg aacgtgatcg gcgagccggt ggacgagctg 300
ggccccatcg agggcgaagc gctgcgcggc atccatcagc cggccccctc ctatgcggag 360
caggccacgg aagctgagat cctcgtcacc ggcatcaagg tggtggatct gctggcgccc 420
tattccaagg gcggcaaggt gggcctgttc ggcggcgccg gcgtgggcaa gaccgtgctc 480
atcatggagc tgatcaacaa cgtggccaag gcgcacggcg gctattccgt gttcgccggc 540
gtgggtgagc gcacccgcga gggcaacgac ctctaccacg agatgatcga gtccaacgtg 600
aacaaggacc cgcacgagaa caatggctcg gcggccggtt ccaagtgcgc cctggtctat 660
ggccagatga acgagccgcc cggcgcccgc gcccgcgtgg ccctcaccgg cctcaccgtc 720
gccgagcatt tccgcgacca gggccaggac gtgctgttct tcgtggacaa catcttccgc 780
ttcacccagg cgggctccga ggtgtcggcg cttctcggcc gcatcccctc ggcggtgggc 840
taccagccga cgctggccac cgacatgggc cagctgcagg agcgcatcac caccaccacc 900
aagggctcca tcacctcggt gcaggccatc tacgtgccgg cggacgatct gaccgatccg 960
gcgccggccg cctccttcgc ccatctggac gccaccacgg tgctgtcgcg ctccatcgcg 1020
gagaagggca tctacccggc ggtggatccg ctggactcca cctcgcgcat gctgtctccc 1080
gccatcctcg gcgacgagca ctacaacacc gcgcgccagg tgcagcagac cctgcagcgc 1140
tacaaggcgc tccaggacat catcgccatc ctgggcatgg acgaactctc cgaagaggac 1200
aagctcaccg tggcccgcgc ccgcaagatc gagcgcttcc tctcccagcc cttccacgtg 1260
gccgaggtgt tcaccggttc gcccggcaag ctggtcgacc tcgccgacac catcaagggc 1320
ttcaagggcc tggtggacgg caagtacgac tacctgcccg agcaggcctt ctacatggtg 1380
ggcaccatcg aagaagccat cgagaagggc aagaagctgg cggccgaggc ggcctga 1437
<210> 33
<211> 478
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 33
Met Ala Asn Lys Val Gly Arg Ile Thr Gln Ile Ile Gly Ala Val Val
1 5 10 15
Asp Val Gln Phe Asp Gly His Leu Pro Ala Ile Leu Asn Ala Ile Glu
20 25 30
Thr Thr Asn Gln Gly Asn Arg Leu Val Leu Glu Val Ala Gln His Leu
35 40 45
Gly Glu Asn Thr Val Arg Cys Ile Ala Met Asp Ala Thr Glu Gly Leu
50 55 60
Val Arg Gly Gln Glu Val Ala Asp Thr Asp Ala Pro Ile Gln Val Pro
65 70 75 80
Val Gly Ala Ala Thr Leu Gly Arg Ile Met Asn Val Ile Gly Glu Pro
85 90 95
Val Asp Glu Leu Gly Pro Ile Glu Gly Glu Ala Leu Arg Gly Ile His
100 105 110
Gln Pro Ala Pro Ser Tyr Ala Glu Gln Ala Thr Glu Ala Glu Ile Leu
115 120 125
Val Thr Gly Ile Lys Val Val Asp Leu Leu Ala Pro Tyr Ser Lys Gly
130 135 140
Gly Lys Val Gly Leu Phe Gly Gly Ala Gly Val Gly Lys Thr Val Leu
145 150 155 160
Ile Met Glu Leu Ile Asn Asn Val Ala Lys Ala His Gly Gly Tyr Ser
165 170 175
Val Phe Ala Gly Val Gly Glu Arg Thr Arg Glu Gly Asn Asp Leu Tyr
180 185 190
His Glu Met Ile Glu Ser Asn Val Asn Lys Asp Pro His Glu Asn Asn
195 200 205
Gly Ser Ala Ala Gly Ser Lys Cys Ala Leu Val Tyr Gly Gln Met Asn
210 215 220
Glu Pro Pro Gly Ala Arg Ala Arg Val Ala Leu Thr Gly Leu Thr Val
225 230 235 240
Ala Glu His Phe Arg Asp Gln Gly Gln Asp Val Leu Phe Phe Val Asp
245 250 255
Asn Ile Phe Arg Phe Thr Gln Ala Gly Ser Glu Val Ser Ala Leu Leu
260 265 270
Gly Arg Ile Pro Ser Ala Val Gly Tyr Gln Pro Thr Leu Ala Thr Asp
275 280 285
Met Gly Gln Leu Gln Glu Arg Ile Thr Thr Thr Thr Lys Gly Ser Ile
290 295 300
Thr Ser Val Gln Ala Ile Tyr Val Pro Ala Asp Asp Leu Thr Asp Pro
305 310 315 320
Ala Pro Ala Ala Ser Phe Ala His Leu Asp Ala Thr Thr Val Leu Ser
325 330 335
Arg Ser Ile Ala Glu Lys Gly Ile Tyr Pro Ala Val Asp Pro Leu Asp
340 345 350
Ser Thr Ser Arg Met Leu Ser Pro Ala Ile Leu Gly Asp Glu His Tyr
355 360 365
Asn Thr Ala Arg Gln Val Gln Gln Thr Leu Gln Arg Tyr Lys Ala Leu
370 375 380
Gln Asp Ile Ile Ala Ile Leu Gly Met Asp Glu Leu Ser Glu Glu Asp
385 390 395 400
Lys Leu Thr Val Ala Arg Ala Arg Lys Ile Glu Arg Phe Leu Ser Gln
405 410 415
Pro Phe His Val Ala Glu Val Phe Thr Gly Ser Pro Gly Lys Leu Val
420 425 430
Asp Leu Ala Asp Thr Ile Lys Gly Phe Lys Gly Leu Val Asp Gly Lys
435 440 445
Tyr Asp Tyr Leu Pro Glu Gln Ala Phe Tyr Met Val Gly Thr Ile Glu
450 455 460
Glu Ala Ile Glu Lys Gly Lys Lys Leu Ala Ala Glu Ala Ala
465 470 475
<210> 34
<211> 876
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 34
atggcgagtc tgaaggacct gagaaaccgc attgcctcgg tgaaggcgac gcagaagatc 60
accaaggcga tgcagatggt cgccgcggcg aagctgcgtc gcgcccaggc ggcggctgaa 120
gcggcccgtc cctatgcgga acgcatggag acggtgctcg gaaatcttgc ctccggcatg 180
gtggtgggcg cgcaggcgcc tgttctcatg accgggacgg gcaagagcga cacccacctg 240
ctgctggtgt gcaccggcga gcgcggcctg tgcggcgcct tcaactcgtc catcgtgcgc 300
ttcgcccgcg agcgggcgca gctgctgctg gccgagggca agaaggtgaa aatcctgtgc 360
gtgggccgca agggccacga gcagctgcgc cgcatctacc cggacaacat catcgacgtg 420
gtggacctgc gcgcggtgcg caacatcggc ttcaaggagg ccgacgccat cgcccgcaag 480
gtgctggccc tgctcgatga aggcgcattc gacgtctgca cgctcttcta ctcccacttc 540
aggagcgtga tcgcccaggt gccgacggcc cagcagctca ttccggccac cttcgacgag 600
cggccggccg tcgccgatgc gccggtctat gaatatgagc cggaggagga ggagatcctc 660
gccgagctgc tgccgcgcaa cgtggcggtg cagatcttca aggccctcct cgagaaccag 720
gcttctttct atggctccca gatgagcgcc atggacaacg ccacgcgcaa tgcgggcgag 780
atgatcaaga agcagacgct cacctacaac cgtacccgcc aggccatgat cacgaaggaa 840
ctcatcgaga tcatctccgg cgccgaggcc gtctga 876
<210> 35
<211> 291
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 35
Met Ala Ser Leu Lys Asp Leu Arg Asn Arg Ile Ala Ser Val Lys Ala
1 5 10 15
Thr Gln Lys Ile Thr Lys Ala Met Gln Met Val Ala Ala Ala Lys Leu
20 25 30
Arg Arg Ala Gln Ala Ala Ala Glu Ala Ala Arg Pro Tyr Ala Glu Arg
35 40 45
Met Glu Thr Val Leu Gly Asn Leu Ala Ser Gly Met Val Val Gly Ala
50 55 60
Gln Ala Pro Val Leu Met Thr Gly Thr Gly Lys Ser Asp Thr His Leu
65 70 75 80
Leu Leu Val Cys Thr Gly Glu Arg Gly Leu Cys Gly Ala Phe Asn Ser
85 90 95
Ser Ile Val Arg Phe Ala Arg Glu Arg Ala Gln Leu Leu Leu Ala Glu
100 105 110
Gly Lys Lys Val Lys Ile Leu Cys Val Gly Arg Lys Gly His Glu Gln
115 120 125
Leu Arg Arg Ile Tyr Pro Asp Asn Ile Ile Asp Val Val Asp Leu Arg
130 135 140
Ala Val Arg Asn Ile Gly Phe Lys Glu Ala Asp Ala Ile Ala Arg Lys
145 150 155 160
Val Leu Ala Leu Leu Asp Glu Gly Ala Phe Asp Val Cys Thr Leu Phe
165 170 175
Tyr Ser His Phe Arg Ser Val Ile Ala Gln Val Pro Thr Ala Gln Gln
180 185 190
Leu Ile Pro Ala Thr Phe Asp Glu Arg Pro Ala Val Ala Asp Ala Pro
195 200 205
Val Tyr Glu Tyr Glu Pro Glu Glu Glu Glu Ile Leu Ala Glu Leu Leu
210 215 220
Pro Arg Asn Val Ala Val Gln Ile Phe Lys Ala Leu Leu Glu Asn Gln
225 230 235 240
Ala Ser Phe Tyr Gly Ser Gln Met Ser Ala Met Asp Asn Ala Thr Arg
245 250 255
Asn Ala Gly Glu Met Ile Lys Lys Gln Thr Leu Thr Tyr Asn Arg Thr
260 265 270
Arg Gln Ala Met Ile Thr Lys Glu Leu Ile Glu Ile Ile Ser Gly Ala
275 280 285
Glu Ala Val
290
<210> 36
<211> 1530
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 36
atggacattc gagccgctga aatctctgcc atcctgaaag agcagatcca gaatttcggc 60
caggaggcgg aagtctccga ggtgggtcag gttctgtccg tgggtgacgg catcgcgcgc 120
gtctacggcc tcgacaacgt ccaggcgggc gagatggtcg agttcgagaa cggcacgcgc 180
ggcatggcgc tgaacctcga gctcgacaat gtcggcatcg tgatcttcgg ttccgaccgc 240
gagatcaagg aaggccagac cgtcaagcgg accggcgcca tcgtggacgc ccccgtcggc 300
aagggcctgc tcggccgcgt cgtggacgct ctcggcaacc cgatcgacgg caagggcccg 360
atcatgttca ccgagcgtcg ccgggtcgac gtgaaggcgc cgggcatcat cccgcgcaag 420
tcggtgcacg agcccatgca gaccggcctg aaggccatcg atgcgctcat ccccatcggc 480
cgcggccagc gcgagctcat catcggcgac cgccagaccg gcaagaccgc cgtggcgctc 540
gactcgatcc tgaaccagaa gcccatcaac cagggcgacg acgagaaggc caagctctac 600
tgcgtctatg tcgcggtggg ccagaagcgt tccactgtcg cgcagttcgt gaaggtgctc 660
gaggagcacg gcgcgctgga atattccatc gtcgtcgccg ccaccgcctc ggacgcggcc 720
cccatgcagt tcctggcgcc gttcaccggc accgccatgg gcgagtattt ccgcgacaac 780
ggcatgcacg ccctcatcat ccatgatgac ctgtccaagc aggccgtggc ctaccgccag 840
atgtcgctgc tgctgcgccg cccgccgggc cgcgaggcct atcccggcga tgtgttctac 900
ctgcactccc gcctcttgga gcgcgccgcc aagctcaatg acgagcacgg cgccggctcg 960
ctgaccgccc tgccggtgat cgagacccag gccaacgacg tgtcggccta catcccgacc 1020
aacgtgatct ccatcaccga cggtcagatc ttccttgaat ccgatctgtt ctaccagggc 1080
atccgcccgg cggtgaacgt gggcctgtcg gtgtcgcgcg tgggctcttc ggcccagatc 1140
aaggcgatga agcaggtggc cggcaagatc aagggcgagc tcgcccagta tcgcgagctg 1200
gcggccttcg cccagttcgg ttcggacctg gacgcggcca cccagaagct gctgaaccgc 1260
ggcgcccgcc tcaccgagct gctgaagcag agccagttct cgcccctcaa ggtggaggag 1320
caggtggcgg tgatctatgc cggcaccaat ggctatctcg atccgctgcc ggtctccaag 1380
gtgcgcgagt tcgagcaggg tctgctcctg tcgctgcgct cgcagcatcc ggagatcctg 1440
gacgccatcc gcacgtccaa ggagctttcc aaggacaccg ccgagaagct gacgaaggcc 1500
atcgacgcct tcgccaagag cttctcctga 1530
<210> 37
<211> 509
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 37
Met Asp Ile Arg Ala Ala Glu Ile Ser Ala Ile Leu Lys Glu Gln Ile
1 5 10 15
Gln Asn Phe Gly Gln Glu Ala Glu Val Ser Glu Val Gly Gln Val Leu
20 25 30
Ser Val Gly Asp Gly Ile Ala Arg Val Tyr Gly Leu Asp Asn Val Gln
35 40 45
Ala Gly Glu Met Val Glu Phe Glu Asn Gly Thr Arg Gly Met Ala Leu
50 55 60
Asn Leu Glu Leu Asp Asn Val Gly Ile Val Ile Phe Gly Ser Asp Arg
65 70 75 80
Glu Ile Lys Glu Gly Gln Thr Val Lys Arg Thr Gly Ala Ile Val Asp
85 90 95
Ala Pro Val Gly Lys Gly Leu Leu Gly Arg Val Val Asp Ala Leu Gly
100 105 110
Asn Pro Ile Asp Gly Lys Gly Pro Ile Met Phe Thr Glu Arg Arg Arg
115 120 125
Val Asp Val Lys Ala Pro Gly Ile Ile Pro Arg Lys Ser Val His Glu
130 135 140
Pro Met Gln Thr Gly Leu Lys Ala Ile Asp Ala Leu Ile Pro Ile Gly
145 150 155 160
Arg Gly Gln Arg Glu Leu Ile Ile Gly Asp Arg Gln Thr Gly Lys Thr
165 170 175
Ala Val Ala Leu Asp Ser Ile Leu Asn Gln Lys Pro Ile Asn Gln Gly
180 185 190
Asp Asp Glu Lys Ala Lys Leu Tyr Cys Val Tyr Val Ala Val Gly Gln
195 200 205
Lys Arg Ser Thr Val Ala Gln Phe Val Lys Val Leu Glu Glu His Gly
210 215 220
Ala Leu Glu Tyr Ser Ile Val Val Ala Ala Thr Ala Ser Asp Ala Ala
225 230 235 240
Pro Met Gln Phe Leu Ala Pro Phe Thr Gly Thr Ala Met Gly Glu Tyr
245 250 255
Phe Arg Asp Asn Gly Met His Ala Leu Ile Ile His Asp Asp Leu Ser
260 265 270
Lys Gln Ala Val Ala Tyr Arg Gln Met Ser Leu Leu Leu Arg Arg Pro
275 280 285
Pro Gly Arg Glu Ala Tyr Pro Gly Asp Val Phe Tyr Leu His Ser Arg
290 295 300
Leu Leu Glu Arg Ala Ala Lys Leu Asn Asp Glu His Gly Ala Gly Ser
305 310 315 320
Leu Thr Ala Leu Pro Val Ile Glu Thr Gln Ala Asn Asp Val Ser Ala
325 330 335
Tyr Ile Pro Thr Asn Val Ile Ser Ile Thr Asp Gly Gln Ile Phe Leu
340 345 350
Glu Ser Asp Leu Phe Tyr Gln Gly Ile Arg Pro Ala Val Asn Val Gly
355 360 365
Leu Ser Val Ser Arg Val Gly Ser Ser Ala Gln Ile Lys Ala Met Lys
370 375 380
Gln Val Ala Gly Lys Ile Lys Gly Glu Leu Ala Gln Tyr Arg Glu Leu
385 390 395 400
Ala Ala Phe Ala Gln Phe Gly Ser Asp Leu Asp Ala Ala Thr Gln Lys
405 410 415
Leu Leu Asn Arg Gly Ala Arg Leu Thr Glu Leu Leu Lys Gln Ser Gln
420 425 430
Phe Ser Pro Leu Lys Val Glu Glu Gln Val Ala Val Ile Tyr Ala Gly
435 440 445
Thr Asn Gly Tyr Leu Asp Pro Leu Pro Val Ser Lys Val Arg Glu Phe
450 455 460
Glu Gln Gly Leu Leu Leu Ser Leu Arg Ser Gln His Pro Glu Ile Leu
465 470 475 480
Asp Ala Ile Arg Thr Ser Lys Glu Leu Ser Lys Asp Thr Ala Glu Lys
485 490 495
Leu Thr Lys Ala Ile Asp Ala Phe Ala Lys Ser Phe Ser
500 505
<210> 38
<211> 555
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 38
gtggcggaaa cgatcgtgtc aggcatggcg ggacgctatg cgaccgcgct gttcgagctg 60
gcggacgaag ccggtgccat cgattccgtc caggcggatc ttgatcgcct gtccggcctt 120
ctggccgaga gcgcggatct ggcgcggctg gtcaagagcc cggtcttcac cgccgagcag 180
cagctcggcg cgatggcggc cattctcgat caagcaggca tttccggcct tgcgggcaaa 240
ttcgtgaagc tggtggcgca gaaccgccgc ctgttcgcac tgccgcgcat gattgccgaa 300
tacgccgtcc tggtggcccg gaagaagggc gagacctcgg cgagcgtgac cgttgccacc 360
cccctgagcg atgagcatct ggccacgctc aaggcggccc tggctgaaaa gaccggcaag 420
gacgtgaagc tcgacgtcac cgtcgatccg tccatcctcg gtggtctcat cgtgaagctc 480
ggctcgcgca tggtcgatgc ttccctgaag accaaactca attctatccg gcatgcgatg 540
aaagaggtcc gctga 555
<210> 39
<211> 184
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 39
Met Ala Glu Thr Ile Val Ser Gly Met Ala Gly Arg Tyr Ala Thr Ala
1 5 10 15
Leu Phe Glu Leu Ala Asp Glu Ala Gly Ala Ile Asp Ser Val Gln Ala
20 25 30
Asp Leu Asp Arg Leu Ser Gly Leu Leu Ala Glu Ser Ala Asp Leu Ala
35 40 45
Arg Leu Val Lys Ser Pro Val Phe Thr Ala Glu Gln Gln Leu Gly Ala
50 55 60
Met Ala Ala Ile Leu Asp Gln Ala Gly Ile Ser Gly Leu Ala Gly Lys
65 70 75 80
Phe Val Lys Leu Val Ala Gln Asn Arg Arg Leu Phe Ala Leu Pro Arg
85 90 95
Met Ile Ala Glu Tyr Ala Val Leu Val Ala Arg Lys Lys Gly Glu Thr
100 105 110
Ser Ala Ser Val Thr Val Ala Thr Pro Leu Ser Asp Glu His Leu Ala
115 120 125
Thr Leu Lys Ala Ala Leu Ala Glu Lys Thr Gly Lys Asp Val Lys Leu
130 135 140
Asp Val Thr Val Asp Pro Ser Ile Leu Gly Gly Leu Ile Val Lys Leu
145 150 155 160
Gly Ser Arg Met Val Asp Ala Ser Leu Lys Thr Lys Leu Asn Ser Ile
165 170 175
Arg His Ala Met Lys Glu Val Arg
180
<210> 40
<211> 492
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 40
atgaccgaaa tggaactggc tgagctctgg gtcgccatcg ccttcctggt tttcgtaggc 60
ctcctgatct atgcgggcgc ccaccgcgcc atcgtctccg ccctggattc ccgcggctcg 120
cgcatcgcct cggaactgga ggaggcccgt cggctcaagg aagaggccca gaagctggtg 180
gccgaattca agcgcaagca gcgcgaggcc gaggccgagg ccgaatccat cgtcaccggc 240
gccaaggccg aggccgagcg cctcgccgcc gaggccaagg cgaagatcga ggatttcgtc 300
acccgccgca ccaagatggc cgaggacaag atcgcccagg ccgagcatca ggctctggcg 360
gacgtgaagt ccatcgccgc cgaggcggcg gccaaggcgg ccgaggtgat cctcggcgcc 420
caggccaccg gcgcggtggc ggagcgtctg ctgtcgggcg ccatctccga ggtcaagacc 480
aagctcaact ga 492
<210> 41
<211> 163
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 41
Met Thr Glu Met Glu Leu Ala Glu Leu Trp Val Ala Ile Ala Phe Leu
1 5 10 15
Val Phe Val Gly Leu Leu Ile Tyr Ala Gly Ala His Arg Ala Ile Val
20 25 30
Ser Ala Leu Asp Ser Arg Gly Ser Arg Ile Ala Ser Glu Leu Glu Glu
35 40 45
Ala Arg Arg Leu Lys Glu Glu Ala Gln Lys Leu Val Ala Glu Phe Lys
50 55 60
Arg Lys Gln Arg Glu Ala Glu Ala Glu Ala Glu Ser Ile Val Thr Gly
65 70 75 80
Ala Lys Ala Glu Ala Glu Arg Leu Ala Ala Glu Ala Lys Ala Lys Ile
85 90 95
Glu Asp Phe Val Thr Arg Arg Thr Lys Met Ala Glu Asp Lys Ile Ala
100 105 110
Gln Ala Glu His Gln Ala Leu Ala Asp Val Lys Ser Ile Ala Ala Glu
115 120 125
Ala Ala Ala Lys Ala Ala Glu Val Ile Leu Gly Ala Gln Ala Thr Gly
130 135 140
Ala Val Ala Glu Arg Leu Leu Ser Gly Ala Ile Ser Glu Val Lys Thr
145 150 155 160
Lys Leu Asn
<210> 42
<211> 729
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 42
atgatgattg catggaagcg gaccttcgca gtcgtgacct tcggggccgc cctgatggcc 60
atgcccgtcg cgggcgtggt cgcagctgag acttctcccg ctccggcggc agtggcgcag 120
gccgatcatg cggtgcccac cgaggcggcc ggccagggca ccgccgatgc ggcccatgcc 180
gccgcgccgg gcgaggccgc ccatggtggc gcggccaagc acgaaaccca tttcccgccc 240
ttcgacggca ccaccttcgc ctcccagttg ctgtggctcg ccgtcacctt cggcctgctt 300
tactacctca tgagcaaggt cacgctgccg cgcatcggcc gcatcctgga agagcgccac 360
gaccgcatcg ccgatgatct ggaggaagcc tccaagcatc gcgccgagag cgaggccgcc 420
cagcgggcct atgagaaggc gctgagcgag gcccgcgcga aggcccattc catcgccgcg 480
gaaacccgcg accgccttgc cgcccacgcc gacaccaacc gcaaggcgct ggagagcgag 540
ctcaccgcca agctgcaggc ggccgaggag cgcatcgcca ccaccaagag cgaagccctc 600
acccatgtgc gcggcatcgc ggtggacgcc acccaatcca tcgtctccac cctcatcggt 660
gtcgcgcccg cggcggccga cgtggaaaaa gcggtggacg gcgccctgtc ccagcacggc 720
caggcctga 729
<210> 43
<211> 242
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 43
Met Met Ile Ala Trp Lys Arg Thr Phe Ala Val Val Thr Phe Gly Ala
1 5 10 15
Ala Leu Met Ala Met Pro Val Ala Gly Val Val Ala Ala Glu Thr Ser
20 25 30
Pro Ala Pro Ala Ala Val Ala Gln Ala Asp His Ala Val Pro Thr Glu
35 40 45
Ala Ala Gly Gln Gly Thr Ala Asp Ala Ala His Ala Ala Ala Pro Gly
50 55 60
Glu Ala Ala His Gly Gly Ala Ala Lys His Glu Thr His Phe Pro Pro
65 70 75 80
Phe Asp Gly Thr Thr Phe Ala Ser Gln Leu Leu Trp Leu Ala Val Thr
85 90 95
Phe Gly Leu Leu Tyr Tyr Leu Met Ser Lys Val Thr Leu Pro Arg Ile
100 105 110
Gly Arg Ile Leu Glu Glu Arg His Asp Arg Ile Ala Asp Asp Leu Glu
115 120 125
Glu Ala Ser Lys His Arg Ala Glu Ser Glu Ala Ala Gln Arg Ala Tyr
130 135 140
Glu Lys Ala Leu Ser Glu Ala Arg Ala Lys Ala His Ser Ile Ala Ala
145 150 155 160
Glu Thr Arg Asp Arg Leu Ala Ala His Ala Asp Thr Asn Arg Lys Ala
165 170 175
Leu Glu Ser Glu Leu Thr Ala Lys Leu Gln Ala Ala Glu Glu Arg Ile
180 185 190
Ala Thr Thr Lys Ser Glu Ala Leu Thr His Val Arg Gly Ile Ala Val
195 200 205
Asp Ala Thr Gln Ser Ile Val Ser Thr Leu Ile Gly Val Ala Pro Ala
210 215 220
Ala Ala Asp Val Glu Lys Ala Val Asp Gly Ala Leu Ser Gln His Gly
225 230 235 240
Gln Ala
<210> 44
<211> 228
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 44
atggaagcgg aagctggaaa gttcatcggt gccggcctcg cctgcctcgg catgggtctc 60
gctggcgtcg gcgtcggtaa catcttcggt aacttcctct ccggcgccct gcgcaacccg 120
tccgctgccg acggccagtt cgcccgcgcc ttcatcggcg ccgccctcgc ggaaggtctc 180
ggcatcttct cgctggtcgt tgcgctcgtc ctgctgttcg tggcctga 228
<210> 45
<211> 75
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 45
Met Glu Ala Glu Ala Gly Lys Phe Ile Gly Ala Gly Leu Ala Cys Leu
1 5 10 15
Gly Met Gly Leu Ala Gly Val Gly Val Gly Asn Ile Phe Gly Asn Phe
20 25 30
Leu Ser Gly Ala Leu Arg Asn Pro Ser Ala Ala Asp Gly Gln Phe Ala
35 40 45
Arg Ala Phe Ile Gly Ala Ala Leu Ala Glu Gly Leu Gly Ile Phe Ser
50 55 60
Leu Val Val Ala Leu Val Leu Leu Phe Val Ala
65 70 75
<210> 46
<211> 753
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 46
atgaccgtcg atccgatcca ccagttcgag atcaagcgct acgtggatct gctgaacgtc 60
ggcggtgtcc agttctcctt caccaacgca acggtgttca tgattggcat cgtcctggtg 120
attttcttct tcctgacttt cgcgacacgc ggtcgcaccc ttgtgccggg ccggatgcag 180
tcggcggcgg agctgagcta cgagttcatc gccaagatgg tgcgcgacgc ggccggcagc 240
gagggaatgg tgttctttcc cttcgtcttc tcgctcttca tgttcgtgct ggtggcgaac 300
gtattggggc tcatccccta caccttcacg gtgaccgccc acctcatcgt caccgccgcc 360
ctggcggcga cggtgatcct caccgtcatc atctacggct tcgtgcggca cggcacccac 420
ttcctgcacc tgttcgtgcc gtcgggcgtg ccgggcttcc tcctgccctt cctcgtggtg 480
atcgaggtgg tgtcgttcct gtcgcggccc atcagcctct cgctgcgtct gttcgccaac 540
atgctggcgg gccacatcgc cctcaaggtg ttcgccttct tcgtcgtggg actggcctcg 600
gccggcgcga tcggctggtt cggcgccacc ctgcccttct tcatgatcgt ggcgctcacc 660
gcgctggagc tgctggtggc ggtgctgcag gcctacgtgt tcgcggtgct gacctcgatc 720
tacctcaacg acgccatcca tcccggccac tga 753
<210> 47
<211> 250
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 47
Met Thr Val Asp Pro Ile His Gln Phe Glu Ile Lys Arg Tyr Val Asp
1 5 10 15
Leu Leu Asn Val Gly Gly Val Gln Phe Ser Phe Thr Asn Ala Thr Val
20 25 30
Phe Met Ile Gly Ile Val Leu Val Ile Phe Phe Phe Leu Thr Phe Ala
35 40 45
Thr Arg Gly Arg Thr Leu Val Pro Gly Arg Met Gln Ser Ala Ala Glu
50 55 60
Leu Ser Tyr Glu Phe Ile Ala Lys Met Val Arg Asp Ala Ala Gly Ser
65 70 75 80
Glu Gly Met Val Phe Phe Pro Phe Val Phe Ser Leu Phe Met Phe Val
85 90 95
Leu Val Ala Asn Val Leu Gly Leu Ile Pro Tyr Thr Phe Thr Val Thr
100 105 110
Ala His Leu Ile Val Thr Ala Ala Leu Ala Ala Thr Val Ile Leu Thr
115 120 125
Val Ile Ile Tyr Gly Phe Val Arg His Gly Thr His Phe Leu His Leu
130 135 140
Phe Val Pro Ser Gly Val Pro Gly Phe Leu Leu Pro Phe Leu Val Val
145 150 155 160
Ile Glu Val Val Ser Phe Leu Ser Arg Pro Ile Ser Leu Ser Leu Arg
165 170 175
Leu Phe Ala Asn Met Leu Ala Gly His Ile Ala Leu Lys Val Phe Ala
180 185 190
Phe Phe Val Val Gly Leu Ala Ser Ala Gly Ala Ile Gly Trp Phe Gly
195 200 205
Ala Thr Leu Pro Phe Phe Met Ile Val Ala Leu Thr Ala Leu Glu Leu
210 215 220
Leu Val Ala Val Leu Gln Ala Tyr Val Phe Ala Val Leu Thr Ser Ile
225 230 235 240
Tyr Leu Asn Asp Ala Ile His Pro Gly His
245 250
<210> 48
<211> 390
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 48
atgtccgagc cgaatgatcc atcccgcagg gacggtgcga aggcgaaaga cgagacgcag 60
gactcccggc ccggtgaggc ggatcttgct cggcgcctcg atgcgctcgg cacctccatc 120
ggtcaggtca agtccagaag cggggagccc gcggcgacgc cgcgcaagga cacctcctcg 180
gcctccggcg cggccctggc gtttcggctg ggcgccgagt ttgtttcagg cgtgctggtg 240
ggctcgctca tcggctacgg gttggattat gcgtttgcga tttcgccctg ggggctgatc 300
gccttcacgc tgatcggctt tgccgccggc gtcctgaaca tgctgcgcgt ggcgaacagc 360
gatgccaagc gccacagcgc ggacaggtga 390
<210> 49
<211> 129
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 49
Met Ser Glu Pro Asn Asp Pro Ser Arg Arg Asp Gly Ala Lys Ala Lys
1 5 10 15
Asp Glu Thr Gln Asp Ser Arg Pro Gly Glu Ala Asp Leu Ala Arg Arg
20 25 30
Leu Asp Ala Leu Gly Thr Ser Ile Gly Gln Val Lys Ser Arg Ser Gly
35 40 45
Glu Pro Ala Ala Thr Pro Arg Lys Asp Thr Ser Ser Ala Ser Gly Ala
50 55 60
Ala Leu Ala Phe Arg Leu Gly Ala Glu Phe Val Ser Gly Val Leu Val
65 70 75 80
Gly Ser Leu Ile Gly Tyr Gly Leu Asp Tyr Ala Phe Ala Ile Ser Pro
85 90 95
Trp Gly Leu Ile Ala Phe Thr Leu Ile Gly Phe Ala Ala Gly Val Leu
100 105 110
Asn Met Leu Arg Val Ala Asn Ser Asp Ala Lys Arg His Ser Ala Asp
115 120 125
Arg
<210> 50
<211> 1929
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 50
atgagttcgc tctccgccac tattcaacag gtcttcaacg agccgggctg cgcgaagaac 60
cagaataagt ccgaggcgga gaagaagaag ggctgcacca agcagctgca acccggcgga 120
gcggccggcg gctgcgcgtt cgacggcgcg aagatcgcgc tccagccctt gaccgacgtc 180
gcccacctgg tgcacggccc catcgcctgc gaaggcaatt cctgggacaa tcgtggcgcc 240
aagtcctccg gctcgaacat ctggcgcacc ggcttcacca cggacatcaa cgaaaccgac 300
gtggtgttcg gcggcgagaa gcgtctgttc aagtccatca aggaaatcat cgagaagtac 360
gacccgccgg ccgtcttcgt ctatcagacc tgcgtccccg ccatgatcgg cgacgacatc 420
gacgcggtgt gcaaggcggc cagggagaag ttcggaaagc cggtgatccc gatcaattcc 480
cccggcttcg tggggccgaa gaatctcggc aacaagctcg ccggcgaggc gctcctcgac 540
catgtgatcg gcaccgagga gcccgattac acgacggcct acgacatcaa catcatcggc 600
gaatacaatc tctccggcga gttgtggcag gtgaagccgc tgctggacga gctgggcatc 660
cgcatcctcg cctgcatctc cggcgacggg aagtacaagg atgtggcgtc ctcccaccgc 720
gccaaggcgg cgatgatggt gtgctccaag gccatgatca acgtggcccg caagatggag 780
gagcgctacg acatcccctt cttcgaaggc tccttctacg gcatcgagga tagctccgat 840
tccctgcgcg agattgcgcg catgctcatc gagaagggcg ccgatccgga gctgatggac 900
cgcaccgagg cgctgattga gcgggaagag aagaaggcgt gggacgccat cgccgcctac 960
aagccccgct tcaaggacaa gaaggtgctg ctcatcaccg gcggcgtgaa atcctggtcg 1020
gtggtggcag cgctccagga agccggcctc gaactggtgg gcacctcggt gaagaagtcc 1080
accaaggagg acaaggagcg catcaaggaa ctgatgggcc aggacgccca catgatcgac 1140
gacatgacgc cccgcgaaat gtacaagatg ctgaaggacg ccaaggcgga catcatgctc 1200
tcgggcgggc gctcgcaatt catcgcgctc aaggccgcca tgccctggct cgacatcaac 1260
caggagcgcc accacgccta tatgggctat gtgggcatgg tgaagctggt cgaggagatc 1320
gacaaggcgc tctacaatcc cgtgtgggaa caggtgcgca agcccgcccc gtgggaaaat 1380
ccggaagaca cctggcaggc ccgtgcgctc gccgaaatgg aggcggaggc cgccgcgctc 1440
gccgccgatc cggtgcgcgc ggaagaggtg cgccggtcca agaagatctg caattgcaag 1500
agcgtcgacc tcggaaccat tgaggacgcc atcaaggctc acgcgctgac caccgtggag 1560
ggtgtgcgag agcacaccaa tgcctcggga ggctgcggag cctgcagcgg gcggatcgag 1620
gagatcttcg aggccgtggg cgttgtcgcc gccccgcctc ccgcggaggc cgccccgtct 1680
ccgcaggaga tcgcgcccga tccgctcgct gcggaggaaa agcgccgcgc caagaaggcc 1740
tgcggctgca aggaggtagc ggtcggcacc attgaggatg ccatccgcgc caagggtctg 1800
cgaaacatcg cggaggtgcg tgcggccacc gatgccaaca ccggctgcgg caattgccag 1860
gagcgggtgg agggcatcct cgaccgggtt ctcgccgagg cggcctcaga actccaggcg 1920
gcggaatag 1929
<210> 51
<211> 642
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 51
Met Ser Ser Leu Ser Ala Thr Ile Gln Gln Val Phe Asn Glu Pro Gly
1 5 10 15
Cys Ala Lys Asn Gln Asn Lys Ser Glu Ala Glu Lys Lys Lys Gly Cys
20 25 30
Thr Lys Gln Leu Gln Pro Gly Gly Ala Ala Gly Gly Cys Ala Phe Asp
35 40 45
Gly Ala Lys Ile Ala Leu Gln Pro Leu Thr Asp Val Ala His Leu Val
50 55 60
His Gly Pro Ile Ala Cys Glu Gly Asn Ser Trp Asp Asn Arg Gly Ala
65 70 75 80
Lys Ser Ser Gly Ser Asn Ile Trp Arg Thr Gly Phe Thr Thr Asp Ile
85 90 95
Asn Glu Thr Asp Val Val Phe Gly Gly Glu Lys Arg Leu Phe Lys Ser
100 105 110
Ile Lys Glu Ile Ile Glu Lys Tyr Asp Pro Pro Ala Val Phe Val Tyr
115 120 125
Gln Thr Cys Val Pro Ala Met Ile Gly Asp Asp Ile Asp Ala Val Cys
130 135 140
Lys Ala Ala Arg Glu Lys Phe Gly Lys Pro Val Ile Pro Ile Asn Ser
145 150 155 160
Pro Gly Phe Val Gly Pro Lys Asn Leu Gly Asn Lys Leu Ala Gly Glu
165 170 175
Ala Leu Leu Asp His Val Ile Gly Thr Glu Glu Pro Asp Tyr Thr Thr
180 185 190
Ala Tyr Asp Ile Asn Ile Ile Gly Glu Tyr Asn Leu Ser Gly Glu Leu
195 200 205
Trp Gln Val Lys Pro Leu Leu Asp Glu Leu Gly Ile Arg Ile Leu Ala
210 215 220
Cys Ile Ser Gly Asp Gly Lys Tyr Lys Asp Val Ala Ser Ser His Arg
225 230 235 240
Ala Lys Ala Ala Met Met Val Cys Ser Lys Ala Met Ile Asn Val Ala
245 250 255
Arg Lys Met Glu Glu Arg Tyr Asp Ile Pro Phe Phe Glu Gly Ser Phe
260 265 270
Tyr Gly Ile Glu Asp Ser Ser Asp Ser Leu Arg Glu Ile Ala Arg Met
275 280 285
Leu Ile Glu Lys Gly Ala Asp Pro Glu Leu Met Asp Arg Thr Glu Ala
290 295 300
Leu Ile Glu Arg Glu Glu Lys Lys Ala Trp Asp Ala Ile Ala Ala Tyr
305 310 315 320
Lys Pro Arg Phe Lys Asp Lys Lys Val Leu Leu Ile Thr Gly Gly Val
325 330 335
Lys Ser Trp Ser Val Val Ala Ala Leu Gln Glu Ala Gly Leu Glu Leu
340 345 350
Val Gly Thr Ser Val Lys Lys Ser Thr Lys Glu Asp Lys Glu Arg Ile
355 360 365
Lys Glu Leu Met Gly Gln Asp Ala His Met Ile Asp Asp Met Thr Pro
370 375 380
Arg Glu Met Tyr Lys Met Leu Lys Asp Ala Lys Ala Asp Ile Met Leu
385 390 395 400
Ser Gly Gly Arg Ser Gln Phe Ile Ala Leu Lys Ala Ala Met Pro Trp
405 410 415
Leu Asp Ile Asn Gln Glu Arg His His Ala Tyr Met Gly Tyr Val Gly
420 425 430
Met Val Lys Leu Val Glu Glu Ile Asp Lys Ala Leu Tyr Asn Pro Val
435 440 445
Trp Glu Gln Val Arg Lys Pro Ala Pro Trp Glu Asn Pro Glu Asp Thr
450 455 460
Trp Gln Ala Arg Ala Leu Ala Glu Met Glu Ala Glu Ala Ala Ala Leu
465 470 475 480
Ala Ala Asp Pro Val Arg Ala Glu Glu Val Arg Arg Ser Lys Lys Ile
485 490 495
Cys Asn Cys Lys Ser Val Asp Leu Gly Thr Ile Glu Asp Ala Ile Lys
500 505 510
Ala His Ala Leu Thr Thr Val Glu Gly Val Arg Glu His Thr Asn Ala
515 520 525
Ser Gly Gly Cys Gly Ala Cys Ser Gly Arg Ile Glu Glu Ile Phe Glu
530 535 540
Ala Val Gly Val Val Ala Ala Pro Pro Pro Ala Glu Ala Ala Pro Ser
545 550 555 560
Pro Gln Glu Ile Ala Pro Asp Pro Leu Ala Ala Glu Glu Lys Arg Arg
565 570 575
Ala Lys Lys Ala Cys Gly Cys Lys Glu Val Ala Val Gly Thr Ile Glu
580 585 590
Asp Ala Ile Arg Ala Lys Gly Leu Arg Asn Ile Ala Glu Val Arg Ala
595 600 605
Ala Thr Asp Ala Asn Thr Gly Cys Gly Asn Cys Gln Glu Arg Val Glu
610 615 620
Gly Ile Leu Asp Arg Val Leu Ala Glu Ala Ala Ser Glu Leu Gln Ala
625 630 635 640
Ala Glu
<210> 52
<211> 1503
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 52
atgagtgtcg cacagtccca gagcgtcgcc gagatcaagg cgcgcaacaa ggaactcatc 60
gaagaggtcc tcaaggtcta tcccgagaag accgccaagc gccgcgccaa gcacctgaac 120
gtccacgaag ccggcaagtc cgactgcggc gtgaagtcca acatcaagtc catcccgggc 180
gtgatgacca tccgcggttg cgcttatgcc ggctccaagg gtgtggtgtg gggtcccatc 240
aaggacatga tccacatctc ccacggcccg gtgggctgcg gccagtatag ctgggccgcc 300
cgccgcaact actatatcgg cacgaccggc atcgacacct tcgtgacgat gcagttcacc 360
tccgacttcc aggagaagga catcgtcttc ggcggcgaca agaagctcgc caagatcatg 420
gacgagatcc aggagctgtt cccgctgaac aacggcatca ccgttcagtc cgagtgcccc 480
atcggcctca tcggcgacga catcgaggcc gtctccaagc agaagtccaa ggagtatgag 540
ggcaagacca tcgtgccggt gcgctgcgag ggcttccgcg gcgtgtccca gtccctgggc 600
caccacatcg ccaacgacgc catccgcgat tgggtgttcg acaagatcgc gcccgacgcc 660
gagccgcgct ttgagccgac cccgtacgac gtcgccatca tcggcgacta caatatcggt 720
ggtgacgcct ggtcgtcccg tatcctcctg gaggagatgg gcctgcgcgt gatcgcccag 780
tggtccggcg acggttcgct cgctgagctg gaggccaccc cgaaggccaa gctcaacgtg 840
ctgcactgct accgctccat gaactacatc tcgcgccaca tggaagagaa gtacggtatc 900
ccgtggtgcg agtacaactt cttcggtcct tccaagatcg ccgagtccct gcgcaagatc 960
gccagctact tcgacgacaa gatcaaggaa ggcgcggagc gcgtcatcgc caagtatcag 1020
ccgctcatgg atgcggtgat cgcgaagtat cgtccccgcc tcgagggcaa gaccgtgatg 1080
ctgtacgtgg gcggcctgcg tccccgtcac gtcatcggcg cctacgagga cctgggcatg 1140
gaagtggtcg gcacgggcta cgagttcgcc cataacgacg actaccagcg caccgcccag 1200
cactacgtca aggatggcac catcatctat gacgacgtga ccggctacga gttcgagaag 1260
ttcgtcgaga agatccagcc ggacctggtc ggttcgggca tcaaggaaaa gtacgtcttc 1320
cagaagatgg gcgtgccgtt ccgccagatg cactcctggg actactcggg cccgtaccac 1380
ggctatgacg gcttcgcgat cttcgcgcgc gacatggaca tggccatcaa cagccccgtg 1440
tggaagatga cccaggctcc gtggaagagc gtccccaagc cgacgatgct cgcggctgaa 1500
tga 1503
<210> 53
<211> 500
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 53
Met Ser Val Ala Gln Ser Gln Ser Val Ala Glu Ile Lys Ala Arg Asn
1 5 10 15
Lys Glu Leu Ile Glu Glu Val Leu Lys Val Tyr Pro Glu Lys Thr Ala
20 25 30
Lys Arg Arg Ala Lys His Leu Asn Val His Glu Ala Gly Lys Ser Asp
35 40 45
Cys Gly Val Lys Ser Asn Ile Lys Ser Ile Pro Gly Val Met Thr Ile
50 55 60
Arg Gly Cys Ala Tyr Ala Gly Ser Lys Gly Val Val Trp Gly Pro Ile
65 70 75 80
Lys Asp Met Ile His Ile Ser His Gly Pro Val Gly Cys Gly Gln Tyr
85 90 95
Ser Trp Ala Ala Arg Arg Asn Tyr Tyr Ile Gly Thr Thr Gly Ile Asp
100 105 110
Thr Phe Val Thr Met Gln Phe Thr Ser Asp Phe Gln Glu Lys Asp Ile
115 120 125
Val Phe Gly Gly Asp Lys Lys Leu Ala Lys Ile Met Asp Glu Ile Gln
130 135 140
Glu Leu Phe Pro Leu Asn Asn Gly Ile Thr Val Gln Ser Glu Cys Pro
145 150 155 160
Ile Gly Leu Ile Gly Asp Asp Ile Glu Ala Val Ser Lys Gln Lys Ser
165 170 175
Lys Glu Tyr Glu Gly Lys Thr Ile Val Pro Val Arg Cys Glu Gly Phe
180 185 190
Arg Gly Val Ser Gln Ser Leu Gly His His Ile Ala Asn Asp Ala Ile
195 200 205
Arg Asp Trp Val Phe Asp Lys Ile Ala Pro Asp Ala Glu Pro Arg Phe
210 215 220
Glu Pro Thr Pro Tyr Asp Val Ala Ile Ile Gly Asp Tyr Asn Ile Gly
225 230 235 240
Gly Asp Ala Trp Ser Ser Arg Ile Leu Leu Glu Glu Met Gly Leu Arg
245 250 255
Val Ile Ala Gln Trp Ser Gly Asp Gly Ser Leu Ala Glu Leu Glu Ala
260 265 270
Thr Pro Lys Ala Lys Leu Asn Val Leu His Cys Tyr Arg Ser Met Asn
275 280 285
Tyr Ile Ser Arg His Met Glu Glu Lys Tyr Gly Ile Pro Trp Cys Glu
290 295 300
Tyr Asn Phe Phe Gly Pro Ser Lys Ile Ala Glu Ser Leu Arg Lys Ile
305 310 315 320
Ala Ser Tyr Phe Asp Asp Lys Ile Lys Glu Gly Ala Glu Arg Val Ile
325 330 335
Ala Lys Tyr Gln Pro Leu Met Asp Ala Val Ile Ala Lys Tyr Arg Pro
340 345 350
Arg Leu Glu Gly Lys Thr Val Met Leu Tyr Val Gly Gly Leu Arg Pro
355 360 365
Arg His Val Ile Gly Ala Tyr Glu Asp Leu Gly Met Glu Val Val Gly
370 375 380
Thr Gly Tyr Glu Phe Ala His Asn Asp Asp Tyr Gln Arg Thr Ala Gln
385 390 395 400
His Tyr Val Lys Asp Gly Thr Ile Ile Tyr Asp Asp Val Thr Gly Tyr
405 410 415
Glu Phe Glu Lys Phe Val Glu Lys Ile Gln Pro Asp Leu Val Gly Ser
420 425 430
Gly Ile Lys Glu Lys Tyr Val Phe Gln Lys Met Gly Val Pro Phe Arg
435 440 445
Gln Met His Ser Trp Asp Tyr Ser Gly Pro Tyr His Gly Tyr Asp Gly
450 455 460
Phe Ala Ile Phe Ala Arg Asp Met Asp Met Ala Ile Asn Ser Pro Val
465 470 475 480
Trp Lys Met Thr Gln Ala Pro Trp Lys Ser Val Pro Lys Pro Thr Met
485 490 495
Leu Ala Ala Glu
500
<210> 54
<211> 1377
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 54
atggccaccg tttccgtctc caagaaggcc tgcgcggtca accccctcaa gatgagccag 60
ccggtgggcg gcgcgctcgc cttcatgggc gtgcgcaagg ccatgccgct gctgcacggc 120
tcgcagggct gcacctcctt cggcctggtg ctgttcgtgc gccacttcaa ggaagccatc 180
cccatgcaga ccaccgccat gagcgaggtg gcgacggttc tgggcggcct tgagaatgtg 240
gagcaggcca ttctcaacat ctacaatcgc accaagccgg agatcatcgg catctgctcc 300
accggcgtca ccgagaccaa gggcgatgat gtcgacggct acatcaagct gatccgggac 360
aagtatcccc agctggccga cttcccgctg gtctatgtct ccacccccga tttcaaggac 420
gccttccagg acggttggga gaagaccgtg gcgaagatgg tggaggcgct ggtgaagccc 480
gccgccgaca agcagaagga caagacccgc gtcaacgtcc tgcccggctg ccacctcacg 540
cccggcgatc tggatgagat gcggaccatc ttcgaggatt tcgggctcac accctatttc 600
ctgccggatc tggccggctc gctggatggg catatccccg aggacttctc gcccaccacc 660
atcggcggca tcggcatcga tgagatcgcc accatgggcg aggcggccca caccatctgc 720
atcggcgcgc agatgcgccg ggcgggcgag gccatggaga agaagaccgg cattcccttc 780
aagctgttcg agcgcctgtg cggcctggag gcgaacgacg ccttcatcat gcacctgtcg 840
cagatctccg gccggccggt gccggtgaag tatcgccggc agcggggcca gctggtggat 900
gccatgctgg acggccactt ccatctgggc ggtcgcaagg tggccatggg ggcggagccg 960
gacctgctct acgacgtggg ctccttcctg cacgagatgg gcgcccacat cctttccgcg 1020
gtcaccacca cccagtcgcc ggtgctggcg cgcctgcctg ccgaggaggt gcttatcggc 1080
gacctggagg atctggagac ccaggcgaag gcgcgcggat gcgatctcct gctcacccat 1140
tcccatgggc gccaggcggc ggagcgcctc cacatcccct tctaccggat cggcattccc 1200
atgtttgacc ggctgggggc ggggcatctg ttgtcggtgg gctatcgcgg cacccgcgac 1260
ctcatcttcc atctcgccaa ccttgtgatc gccgaccacg aggaaaatca cgagccgacg 1320
cccgacacct gggccaccgg ccatggcgag catgccgccg cccccacttc ccattga 1377
<210> 55
<211> 458
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 55
Met Ala Thr Val Ser Val Ser Lys Lys Ala Cys Ala Val Asn Pro Leu
1 5 10 15
Lys Met Ser Gln Pro Val Gly Gly Ala Leu Ala Phe Met Gly Val Arg
20 25 30
Lys Ala Met Pro Leu Leu His Gly Ser Gln Gly Cys Thr Ser Phe Gly
35 40 45
Leu Val Leu Phe Val Arg His Phe Lys Glu Ala Ile Pro Met Gln Thr
50 55 60
Thr Ala Met Ser Glu Val Ala Thr Val Leu Gly Gly Leu Glu Asn Val
65 70 75 80
Glu Gln Ala Ile Leu Asn Ile Tyr Asn Arg Thr Lys Pro Glu Ile Ile
85 90 95
Gly Ile Cys Ser Thr Gly Val Thr Glu Thr Lys Gly Asp Asp Val Asp
100 105 110
Gly Tyr Ile Lys Leu Ile Arg Asp Lys Tyr Pro Gln Leu Ala Asp Phe
115 120 125
Pro Leu Val Tyr Val Ser Thr Pro Asp Phe Lys Asp Ala Phe Gln Asp
130 135 140
Gly Trp Glu Lys Thr Val Ala Lys Met Val Glu Ala Leu Val Lys Pro
145 150 155 160
Ala Ala Asp Lys Gln Lys Asp Lys Thr Arg Val Asn Val Leu Pro Gly
165 170 175
Cys His Leu Thr Pro Gly Asp Leu Asp Glu Met Arg Thr Ile Phe Glu
180 185 190
Asp Phe Gly Leu Thr Pro Tyr Phe Leu Pro Asp Leu Ala Gly Ser Leu
195 200 205
Asp Gly His Ile Pro Glu Asp Phe Ser Pro Thr Thr Ile Gly Gly Ile
210 215 220
Gly Ile Asp Glu Ile Ala Thr Met Gly Glu Ala Ala His Thr Ile Cys
225 230 235 240
Ile Gly Ala Gln Met Arg Arg Ala Gly Glu Ala Met Glu Lys Lys Thr
245 250 255
Gly Ile Pro Phe Lys Leu Phe Glu Arg Leu Cys Gly Leu Glu Ala Asn
260 265 270
Asp Ala Phe Ile Met His Leu Ser Gln Ile Ser Gly Arg Pro Val Pro
275 280 285
Val Lys Tyr Arg Arg Gln Arg Gly Gln Leu Val Asp Ala Met Leu Asp
290 295 300
Gly His Phe His Leu Gly Gly Arg Lys Val Ala Met Gly Ala Glu Pro
305 310 315 320
Asp Leu Leu Tyr Asp Val Gly Ser Phe Leu His Glu Met Gly Ala His
325 330 335
Ile Leu Ser Ala Val Thr Thr Thr Gln Ser Pro Val Leu Ala Arg Leu
340 345 350
Pro Ala Glu Glu Val Leu Ile Gly Asp Leu Glu Asp Leu Glu Thr Gln
355 360 365
Ala Lys Ala Arg Gly Cys Asp Leu Leu Leu Thr His Ser His Gly Arg
370 375 380
Gln Ala Ala Glu Arg Leu His Ile Pro Phe Tyr Arg Ile Gly Ile Pro
385 390 395 400
Met Phe Asp Arg Leu Gly Ala Gly His Leu Leu Ser Val Gly Tyr Arg
405 410 415
Gly Thr Arg Asp Leu Ile Phe His Leu Ala Asn Leu Val Ile Ala Asp
420 425 430
His Glu Glu Asn His Glu Pro Thr Pro Asp Thr Trp Ala Thr Gly His
435 440 445
Gly Glu His Ala Ala Ala Pro Thr Ser His
450 455
<210> 56
<211> 1560
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 56
atgccacaaa atgctgacaa tgtgctcgat cacttcgagc tcttccgtgg tcccgaatac 60
cagcagatgc tggccaataa gaaaaagatg ttcgagaacc cccgcgatcc ggccgaagtc 120
gagcgcgtgc gggaatgggc gaagactcct gaatacaagg agctgaactt cgcccgcgag 180
gcgctcaccg tgaatccggc caaggcttgt cagccgctgg gcgcggtgtt cgtcgccgtc 240
ggcttcgaga gcacgatccc cttcgtgcac ggctcgcagg gttgcgtcgc gtattaccgc 300
tcgcacctct cccgccactt caaggagccg tcctcctgcg tctcctcgtc catgaccgag 360
gatgcggcgg tgttcggcgg cctcaacaac atgattgacg gcctcgccaa cacctacaac 420
atgtacaagc cgaagatgat cgccgtctcc accacctgca tggcggaagt catcggcgac 480
gatctgaacg ccttcatcaa gaccgcgaag gaaaagggct cggttccggc cgaatacgac 540
gtgcccttcg cccacacccc ggcgttcgtc ggcagccatg tcaccggcta cgacaatgcg 600
ctcaagggca tcctcgagca cttctgggac ggcaaggccg gcaccgcgcc gaagctggag 660
cgcgttccca acgagaagat caacttcatc ggcggcttcg acggctacac cgtcggcaac 720
actcgcgaag tgaagcgcat cttcgaggcg ttcggcgccg attacaccat cctcgccgac 780
aattccgaag tgttcgacac cccgaccgac ggcgagttcc gcatgtatga cggcggcacg 840
accctggagg acgcggcgaa cgcggtgcac gccaaggcca ccatctccat gcaggaatac 900
tgcacggaga agaccctgcc catgatcgcc ggtcatggcc aggacgtggt cgccctcaac 960
caccccgtgg gcgtgggcgg caccgacaag ttcctcatgg agatcgcccg cctcaccggc 1020
aaggagatcc ccgaggagct gacccgcgag cgcggccgtc tcgtggacgc tatcgcggac 1080
tcttccgcgc acatccacgg caagaagttc gccatctacg gcgatccgga tctgtgcctg 1140
ggcctcgccg cgttcctgct ggagctgggc gccgagccga cccatgtgct ggccaccaac 1200
ggcaccaaga agtgggccga gaaggttcag gaactgttcg actcttcgcc gttcggcgcc 1260
aactgcaagg tctatcccgg caaggacctg tggcacatgc gctcgctcct gttcgtggag 1320
ccggtggatt tcatcatcgg caacacctac ggcaagtatc tcgagcgcga cacgggcacc 1380
ccgctgatcc gtatcggctt cccggtgttc gaccgtcacc accaccaccg ccgtccggtg 1440
tggggctatc agggcggcat gaacgtcctg atcacgatcc tcgacaagat ctttgacgag 1500
atcgaccgca acaccaacgt gccggccaag accgactact cgttcgacat cattcgttga 1560
<210> 57
<211> 519
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 57
Met Pro Gln Asn Ala Asp Asn Val Leu Asp His Phe Glu Leu Phe Arg
1 5 10 15
Gly Pro Glu Tyr Gln Gln Met Leu Ala Asn Lys Lys Lys Met Phe Glu
20 25 30
Asn Pro Arg Asp Pro Ala Glu Val Glu Arg Val Arg Glu Trp Ala Lys
35 40 45
Thr Pro Glu Tyr Lys Glu Leu Asn Phe Ala Arg Glu Ala Leu Thr Val
50 55 60
Asn Pro Ala Lys Ala Cys Gln Pro Leu Gly Ala Val Phe Val Ala Val
65 70 75 80
Gly Phe Glu Ser Thr Ile Pro Phe Val His Gly Ser Gln Gly Cys Val
85 90 95
Ala Tyr Tyr Arg Ser His Leu Ser Arg His Phe Lys Glu Pro Ser Ser
100 105 110
Cys Val Ser Ser Ser Met Thr Glu Asp Ala Ala Val Phe Gly Gly Leu
115 120 125
Asn Asn Met Ile Asp Gly Leu Ala Asn Thr Tyr Asn Met Tyr Lys Pro
130 135 140
Lys Met Ile Ala Val Ser Thr Thr Cys Met Ala Glu Val Ile Gly Asp
145 150 155 160
Asp Leu Asn Ala Phe Ile Lys Thr Ala Lys Glu Lys Gly Ser Val Pro
165 170 175
Ala Glu Tyr Asp Val Pro Phe Ala His Thr Pro Ala Phe Val Gly Ser
180 185 190
His Val Thr Gly Tyr Asp Asn Ala Leu Lys Gly Ile Leu Glu His Phe
195 200 205
Trp Asp Gly Lys Ala Gly Thr Ala Pro Lys Leu Glu Arg Val Pro Asn
210 215 220
Glu Lys Ile Asn Phe Ile Gly Gly Phe Asp Gly Tyr Thr Val Gly Asn
225 230 235 240
Thr Arg Glu Val Lys Arg Ile Phe Glu Ala Phe Gly Ala Asp Tyr Thr
245 250 255
Ile Leu Ala Asp Asn Ser Glu Val Phe Asp Thr Pro Thr Asp Gly Glu
260 265 270
Phe Arg Met Tyr Asp Gly Gly Thr Thr Leu Glu Asp Ala Ala Asn Ala
275 280 285
Val His Ala Lys Ala Thr Ile Ser Met Gln Glu Tyr Cys Thr Glu Lys
290 295 300
Thr Leu Pro Met Ile Ala Gly His Gly Gln Asp Val Val Ala Leu Asn
305 310 315 320
His Pro Val Gly Val Gly Gly Thr Asp Lys Phe Leu Met Glu Ile Ala
325 330 335
Arg Leu Thr Gly Lys Glu Ile Pro Glu Glu Leu Thr Arg Glu Arg Gly
340 345 350
Arg Leu Val Asp Ala Ile Ala Asp Ser Ser Ala His Ile His Gly Lys
355 360 365
Lys Phe Ala Ile Tyr Gly Asp Pro Asp Leu Cys Leu Gly Leu Ala Ala
370 375 380
Phe Leu Leu Glu Leu Gly Ala Glu Pro Thr His Val Leu Ala Thr Asn
385 390 395 400
Gly Thr Lys Lys Trp Ala Glu Lys Val Gln Glu Leu Phe Asp Ser Ser
405 410 415
Pro Phe Gly Ala Asn Cys Lys Val Tyr Pro Gly Lys Asp Leu Trp His
420 425 430
Met Arg Ser Leu Leu Phe Val Glu Pro Val Asp Phe Ile Ile Gly Asn
435 440 445
Thr Tyr Gly Lys Tyr Leu Glu Arg Asp Thr Gly Thr Pro Leu Ile Arg
450 455 460
Ile Gly Phe Pro Val Phe Asp Arg His His His His Arg Arg Pro Val
465 470 475 480
Trp Gly Tyr Gln Gly Gly Met Asn Val Leu Ile Thr Ile Leu Asp Lys
485 490 495
Ile Phe Asp Glu Ile Asp Arg Asn Thr Asn Val Pro Ala Lys Thr Asp
500 505 510
Tyr Ser Phe Asp Ile Ile Arg
515
<210> 58
<211> 621
<212> DNA
<213> Unknown
<220>
<223> novel strain
<400> 58
gtggagtccg gtggtcctga gccgggcgtg ggctgcgccg gccgcggcgt gatcacctcc 60
atcaacttcc tggaggagaa cggcgcctac gaggacatcg actatgtgtc ctacgacgtg 120
ctgggcgacg tggtgtgcgg cggcttcgcc atgcccatcc gcgagaacaa ggcgcaggaa 180
atctacatcg tgatgtccgg cgagatgatg gccatgtatg cggccaacaa catctccaag 240
ggcatcctga agtatgccaa ttccggcggc gtgcgcctgg gcgggctggt ctgcaacgag 300
cgccagaccg acaaggagct ggagctggcg gaggctctgg cgaagaagct cggcaccgag 360
ctgatctact tcgtgccgcg cgacaacatc gtgcagcatg ccgagctgcg ccgcatgaca 420
gtgatcgagt atgcgcccga ttccgcccag gcccagcact accggaacct ggccgagaag 480
gtgcacgcca acaagggcaa cggcatcatc ccgaccccga tcaccatgga cgagctggaa 540
gacatgctca tggagcacgg catcatgaag gccgtggacg agagccagat cggcaagacc 600
gccgccgagc tcgccgtctg a 621
<210> 59
<211> 206
<212> PRT
<213> Unknown
<220>
<223> novel strain
<400> 59
Met Glu Ser Gly Gly Pro Glu Pro Gly Val Gly Cys Ala Gly Arg Gly
1 5 10 15
Val Ile Thr Ser Ile Asn Phe Leu Glu Glu Asn Gly Ala Tyr Glu Asp
20 25 30
Ile Asp Tyr Val Ser Tyr Asp Val Leu Gly Asp Val Val Cys Gly Gly
35 40 45
Phe Ala Met Pro Ile Arg Glu Asn Lys Ala Gln Glu Ile Tyr Ile Val
50 55 60
Met Ser Gly Glu Met Met Ala Met Tyr Ala Ala Asn Asn Ile Ser Lys
65 70 75 80
Gly Ile Leu Lys Tyr Ala Asn Ser Gly Gly Val Arg Leu Gly Gly Leu
85 90 95
Val Cys Asn Glu Arg Gln Thr Asp Lys Glu Leu Glu Leu Ala Glu Ala
100 105 110
Leu Ala Lys Lys Leu Gly Thr Glu Leu Ile Tyr Phe Val Pro Arg Asp
115 120 125
Asn Ile Val Gln His Ala Glu Leu Arg Arg Met Thr Val Ile Glu Tyr
130 135 140
Ala Pro Asp Ser Ala Gln Ala Gln His Tyr Arg Asn Leu Ala Glu Lys
145 150 155 160
Val His Ala Asn Lys Gly Asn Gly Ile Ile Pro Thr Pro Ile Thr Met
165 170 175
Asp Glu Leu Glu Asp Met Leu Met Glu His Gly Ile Met Lys Ala Val
180 185 190
Asp Glu Ser Gln Ile Gly Lys Thr Ala Ala Glu Leu Ala Val
195 200 205
Claims (20)
- 단리된 박테리아 균주 VTT-E-193585 또는 이의 유도체로서, 상기 유도체는 바람직하게는 에너지 공급원으로서 수소 기체 및 유일한 탄소 공급원으로서 이산화탄소를 사용하여 성장하는 능력을 보유한 것인 단리된 박테리아 균주 VTT-E-193585 또는 이의 유도체.
- 청구항 1에 따른 박테리아 균주를 포함하는 배양물.
- 바이오매스를 생산하는 방법으로서, 상기 방법은 청구항 1의 박테리아 균주를 배양하는 단계를 포함하는 바이오매스를 생산하는 방법.
- 청구항 3에 있어서, 에너지 공급원으로서 수소 및 무기 탄소 공급원을 사용하여 연속 배양으로 박테리아 균주를 배양하는 단계를 포함하고, 상기 무기 탄소 공급원은 이산화탄소를 포함하는 것인 방법.
- 바이오매스를 생산하는 방법으로서, 상기 방법은 에너지 공급원으로서 수소 및 무기 탄소 공급원을 사용하여 연속 배양으로 크산토박터 (Xanthobacter) 속의 박테리아 균주를 배양하는 단계를 포함하고, 상기 무기 탄소 공급원은 이산화탄소를 포함하는 것인 방법.
- 청구항 4 또는 5에 있어서, 배양물 중 용존 산소는 5% 내지 10%로 유지되는 것인 방법.
- 청구항 4 내지 6 중 어느 한 항에 있어서, 암모늄, 우레아, 니트레이트 및/또는 질소 기체가 질소 공급원으로서 사용되는 것인 방법.
- 청구항 4 내지 7 중 어느 한 항에 있어서, 배양 배지는 미네랄을 포함하고, 상기 미네랄은 1 g/L 미만의 클로라이드 염 (chloride salts), 예컨대 0.25 g/L 미만의 클로라이드 염, 예를 들어 0.1 g/l 미만의 클로라이드 염을 포함하며, 바람직하게는 클로라이드 염을 포함하지 않는 것인 방법.
- 청구항 4 내지 8 중 어느 한 항에 있어서, 배양 배지는 비타민을 함유하지 않는 것인 방법.
- 청구항 4 내지 9 중 어느 한 항에 있어서, 배양물의 pH는 5.5 내지 8.0, 예를 들어 6.5 내지 7.0, 예컨대 6.8로 유지되는 것인 방법.
- 청구항 4 내지 10 중 어느 한 항에 있어서, 상기 배양물은 25℃ 내지 40℃, 예를 들어 28℃ 내지 32℃, 예컨대 30℃의 온도에서 성장되는 것인 방법.
- 청구항 4 내지 11 중 어느 한 항에 있어서, 박테리아 균주는 0.04-0.12 h-1의 성장 속도를 갖는 것인 방법.
- 청구항 4 내지 12 중 어느 한 항에 있어서, 박테리아 균주는 VTT-E-193585 또는 X. 타게티디스 (X. tagetidis)인 것인 방법.
- 청구항 4 내지 13 중 어느 한 항에 있어서, 박테리아 균주는 서열번호: 1에 제시된 16S 리보솜 RNA, 또는 서열번호: 1과 최대 20개, 예를 들어 1 내지 10개, 예컨대 1 내지 5개의 뉴클레오티드 차이, 예를 들어 서열번호: 1과 1개, 2개 또는 3개의 뉴클레오티드 차이를 갖는 16S 리보솜 RNA를 포함하는 것인 방법.
- 청구항 4 내지 14 중 어느 한 항에 있어서, 박테리아 균주는:
i) 서열번호: 3에 제시된 서열, 또는 서열번호: 3에 제시된 서열에 대해 93% 초과의 동일성, 예를 들어 95% 초과의 동일성, 예컨대 96% 초과의 동일성, 예를 들어 97% 초과의 동일성, 예컨대 98% 초과의 동일성, 예를 들어 99% 초과의 서열 동일성을 갖는 서열을 가진 리불로스-1,5-비스포스페이트 카복실라제/옥시게나제 (rubisco) 큰 사슬을 코딩하는 유전자, 및/또는
ii) 서열번호: 5에 제시된 서열, 또는 서열번호: 5에 제시된 서열에 대해 83% 초과의 서열 동일성, 예를 들어 86% 초과의 동일성, 예컨대 90% 초과의 동일성, 예를 들어 95% 초과의 동일성, 예컨대 96% 초과의 동일성, 예를 들어 97% 초과의 동일성, 예컨대 98% 초과의 동일성, 예를 들어 99% 초과의 서열 동일성을 갖는 서열을 가진 리불로스-1,5-비스포스페이트 카복실라제/옥시게나제 (rubisco) 작은 사슬을 코딩하는 유전자, 및/또는
iii) 서열번호: 7에 제시된 서열, 또는 서열번호: 7에 제시된 서열에 대해 70% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 알파를 코딩하는 유전자, 및/또는
iv) 서열번호: 9에 제시된 서열, 또는 서열번호: 9에 제시된 서열에 대해 77% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 베타를 코딩하는 유전자, 및/또는
v) 서열번호: 11에 제시된 서열, 또는 서열번호: 11에 제시된 서열에 대해 70% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 감마를 코딩하는 유전자, 및/또는
vi) 서열번호: 13에 제시된 서열, 또는 서열번호: 13에 제시된 서열에 대해 79% 초과의 서열 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NAD+-환원 하이드로게나제 HoxS 서브유닛 델타를 코딩하는 유전자, 및/또는
vii) 서열번호: 15에 제시된 서열, 또는 서열번호: 15에 제시된 서열에 대해 84% 초과의 서열 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NiFeSe 하이드로게나제 큰 서브유닛을 코딩하는 유전자, 및/또는
viii) 서열번호: 17에 제시된 서열, 또는 서열번호: 17에 제시된 서열에 대해 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 NiFeSe 하이드로게나제 작은 서브유닛을 코딩하는 유전자, 및/또는
ix) 서열번호: 19에 제시된 서열, 또는 서열번호: 19에 제시된 서열에 대해 70% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 감마 사슬 atpG_1을 코딩하는 유전자, 및/또는
x) 서열번호: 21에 제시된 서열, 또는 서열번호: 21에 제시된 서열에 대해 78% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 알파 atpA_1을 코딩하는 유전자, 및/또는
xi) 서열번호: 23에 제시된 서열, 또는 서열번호: 23에 제시된 서열에 대해 62% 초과의 동일성, 예를 들어 70% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 b atpF_1을 코딩하는 유전자, 및/또는
xii) 서열번호: 25에 제시된 서열, 또는 서열번호: 25에 제시된 서열에 대해 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 c, 나트륨 이온 특이적 atpE_1을 코딩하는 유전자, 및/또는
xiii) 서열번호: 27에 제시된 서열, 또는 서열번호: 27에 제시된 서열에 대해 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 a atpB_1을 코딩하는 유전자, 및/또는
xiv) 서열번호: 29에 제시된 서열, 또는 서열번호: 29에 제시된 서열에 대해 71% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 엡실론 사슬 atpC_1을 코딩하는 유전자, 및/또는
xv) 서열번호: 31에 제시된 서열, 또는 서열번호: 31에 제시된 서열에 대해 84% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 베타 atpD_1을 코딩하는 유전자, 및/또는
xvi) 서열번호: 33에 제시된 서열, 또는 서열번호: 33에 제시된 서열에 대해 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 베타 atpD_2를 코딩하는 유전자, 및/또는
xvii) 서열번호: 35에 제시된 서열, 또는 서열번호: 35에 제시된 서열에 대해 86% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 감마 사슬 atpG_2를 코딩하는 유전자, 및/또는
xviii) 서열번호: 37에 제시된 서열, 또는 서열번호: 37에 제시된 서열에 대해 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 알파 atpA_2를 코딩하는 유전자, 및/또는
xix) 서열번호: 39에 제시된 서열, 또는 서열번호: 39에 제시된 서열에 대해 85% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 델타 atpH를 코딩하는 유전자, 및/또는
xx) 서열번호: 41에 제시된 서열, 또는 서열번호: 41에 제시된 서열에 대해 87% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 b atpF_2를 코딩하는 유전자, 및/또는
xxi) 서열번호: 43에 제시된 서열, 또는 서열번호: 43에 제시된 서열에 대해 81% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 b' atpG_3을 코딩하는 유전자, 및/또는
xxii) 서열번호: 45에 제시된 서열, 또는 서열번호: 45에 제시된 서열에 대해 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 c atpE_2를 코딩하는 유전자, 및/또는
xxiii) 서열번호: 47에 제시된 서열, 또는 서열번호: 47에 제시된 서열에 대해 92% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 서브유닛 a atpB_2를 코딩하는 유전자, 및/또는
xiv) 서열번호: 49에 제시된 서열, 또는 서열번호: 49에 제시된 서열에 대해 60% 초과의 동일성, 예를 들어 70% 초과의 동일성, 예컨대 80% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 ATP 신타제 단백질 I atpI를 코딩하는 유전자, 및/또는
xv) 서열번호: 51에 제시된 서열, 또는 서열번호: 51에 제시된 서열에 대해 60% 초과의 동일성, 예를 들어 70% 초과의 동일성, 예컨대 92% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_1을 코딩하는 유전자, 및/또는
xvi) 서열번호: 53에 제시된 서열, 또는 서열번호: 53에 제시된 서열에 대해 60% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 알파 사슬 nifD_2를 코딩하는 유전자, 및/또는
xvii) 서열번호: 55에 제시된 서열, 또는 서열번호: 55에 제시된 서열에 대해 87% 초과의 동일성, 예를 들어 90% 초과의 동일성, 예컨대 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_1을 코딩하는 유전자, 및/또는
xviii) 서열번호: 57에 제시된 서열, 또는 서열번호: 57에 제시된 서열에 대해 95% 초과의 동일성, 예를 들어 96% 초과의 동일성, 예컨대 97% 초과의 동일성, 예를 들어 98% 초과의 동일성, 예컨대 99% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 몰리브덴-철 단백질 베타 사슬 nifK_2를 코딩하는 유전자, 및/또는
xxix) 서열번호: 59에 제시된 서열, 또는 서열번호: 59에 제시된 서열에 대해 98.5% 초과의 서열 동일성을 갖는 서열을 가진 니트로게나제 철 단백질 nifH를 코딩하는 유전자를 포함하고, 상기 균주는 바람직하게는:
iii), iv), v) 및 vi)에 기재된 유전자들,
vii) 및 viii)에 기재된 유전자들, 또는
iii), iv), v), vi), vii) 및 viii)에 기재된 유전자들을 포함하는 것인 방법. - 청구항 4 내지 15 중 어느 한 항에 있어서, 배양 중에 생산된 바이오매스를 수확하는 추가 단계를 포함하고, 선택적으로 상기 바이오매스를 건조시키는 추가 단계를 포함하는 것인 방법.
- 단백질을 생산하는 방법으로서, 청구항 16에 따른 방법을 수행하는 단계 및 상기 바이오매스로부터 단백질을 단리하는 추가 단계를 포함하고, 상기 방법으로 단백질 분획 및 비-단백질 성분을 포함하는 분획을 수득하는 것인 방법.
- 청구항 16 또는 17에 있어서, 상기 바이오매스, 상기 단백질 분획 또는 상기 비-단백질 성분을 포함하는 분획으로부터 식품 또는 사료 제품을 제조하는 추가 단계를 포함하는 것인 방법.
- 청구항 4 내지 18 중 어느 한 항에 따른 방법에 의해 수득되거나 또는 수득 가능한 바이오매스, 단백질 또는 비-단백질 성분과 같은 산물.
- 청구항 18에 따른 방법에 의해 수득되거나 또는 수득 가능한 식품 또는 사료 제품.
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